CRISPR/Cas system-guided plasmid mutagenesis without sequence restriction Article Swipe
Fengjiao Zhao
,
Feng Chen
,
Huahang Yu
,
Siyue Fan
,
Bai Min
,
Jing Xue
,
Yue Zhao
,
Xiaolei Zuo
,
Chunhai Fan
,
Yongxi Zhao
·
YOU?
·
· 2022
· Open Access
·
· DOI: https://doi.org/10.1016/j.fmre.2022.06.017
YOU?
·
· 2022
· Open Access
·
· DOI: https://doi.org/10.1016/j.fmre.2022.06.017
Plasmid mutagenesis is an essential step to engineer protein variants with desired properties. Direct generation of mutations on plasmids is a promising method compared to PCR-based methods and subcloning, yet it suffers from the sequence restriction in the generation of single-stranded circular plasmids. Here we demonstrate the CRISPR/Cas system-guided plasmid mutagenesis, a method using gRNA/Cas9 nickase complex to generate single-stranded circular plasmids as polymerization templates for mutagenesis. This method can directly create user-defined mutation libraries on the plasmid. It offers broad sequence programmability and even covers methylated plasmids. We further combine this method with rational design to engineer genome-editing protein FnCpf1. The FnCpf1 variants with mutations in the PAM-binding groove were generated. Several variants expanded the PAM range and exhibited lower off-target effect, which can loosen PAM constraint and enhance target specificity for genome editing. This work presents an effective tool for plasmid mutagenesis and protein engineering studies.
Related Topics
Concepts
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1016/j.fmre.2022.06.017
- OA Status
- gold
- References
- 46
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4285500066
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
-
https://openalex.org/W4285500066Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.1016/j.fmre.2022.06.017Digital Object Identifier
- Title
-
CRISPR/Cas system-guided plasmid mutagenesis without sequence restrictionWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2022Year of publication
- Publication date
-
2022-07-15Full publication date if available
- Authors
-
Fengjiao Zhao, Feng Chen, Huahang Yu, Siyue Fan, Bai Min, Jing Xue, Yue Zhao, Xiaolei Zuo, Chunhai Fan, Yongxi ZhaoList of authors in order
- Landing page
-
https://doi.org/10.1016/j.fmre.2022.06.017Publisher landing page
- Open access
-
YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
- OA URL
-
https://doi.org/10.1016/j.fmre.2022.06.017Direct OA link when available
- Concepts
-
Plasmid, CRISPR, Mutagenesis, Cas9, Biology, Computational biology, Genetics, Genome editing, Subcloning, DNA, Mutation, GeneTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
0Total citation count in OpenAlex
- References (count)
-
46Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
Full payload
| id | https://openalex.org/W4285500066 |
|---|---|
| doi | https://doi.org/10.1016/j.fmre.2022.06.017 |
| ids.doi | https://doi.org/10.1016/j.fmre.2022.06.017 |
| ids.pmid | https://pubmed.ncbi.nlm.nih.gov/40777797 |
| ids.openalex | https://openalex.org/W4285500066 |
| fwci | 0.0 |
| type | article |
| title | CRISPR/Cas system-guided plasmid mutagenesis without sequence restriction |
| awards[0].id | https://openalex.org/G1552126911 |
| awards[0].funder_id | https://openalex.org/F4320321543 |
| awards[0].display_name | |
| awards[0].funder_award_id | 2018M643614 |
| awards[0].funder_display_name | China Postdoctoral Science Foundation |
| awards[1].id | https://openalex.org/G5733994830 |
| awards[1].funder_id | https://openalex.org/F4320336567 |
| awards[1].display_name | |
| awards[1].funder_award_id | 2021JQ-016 |
| awards[1].funder_display_name | Natural Science Basic Research Program of Shaanxi Province |
| awards[2].id | https://openalex.org/G1157647398 |
| awards[2].funder_id | https://openalex.org/F4320321001 |
| awards[2].display_name | |
| awards[2].funder_award_id | 22125404 |
| awards[2].funder_display_name | National Natural Science Foundation of China |
| awards[3].id | https://openalex.org/G8784620080 |
| awards[3].funder_id | https://openalex.org/F4320321001 |
| awards[3].display_name | |
| awards[3].funder_award_id | 92068118 |
| awards[3].funder_display_name | National Natural Science Foundation of China |
| awards[4].id | https://openalex.org/G5983096502 |
| awards[4].funder_id | https://openalex.org/F4320321001 |
| awards[4].display_name | |
| awards[4].funder_award_id | 32101013 |
| awards[4].funder_display_name | National Natural Science Foundation of China |
| biblio.issue | 4 |
| biblio.volume | 5 |
| biblio.last_page | 1487 |
| biblio.first_page | 1481 |
| topics[0].id | https://openalex.org/T10878 |
| topics[0].field.id | https://openalex.org/fields/13 |
| topics[0].field.display_name | Biochemistry, Genetics and Molecular Biology |
| topics[0].score | 1.0 |
| topics[0].domain.id | https://openalex.org/domains/1 |
| topics[0].domain.display_name | Life Sciences |
| topics[0].subfield.id | https://openalex.org/subfields/1312 |
| topics[0].subfield.display_name | Molecular Biology |
| topics[0].display_name | CRISPR and Genetic Engineering |
| topics[1].id | https://openalex.org/T10207 |
| topics[1].field.id | https://openalex.org/fields/13 |
| topics[1].field.display_name | Biochemistry, Genetics and Molecular Biology |
| topics[1].score | 0.9876000285148621 |
| topics[1].domain.id | https://openalex.org/domains/1 |
| topics[1].domain.display_name | Life Sciences |
| topics[1].subfield.id | https://openalex.org/subfields/1312 |
| topics[1].subfield.display_name | Molecular Biology |
| topics[1].display_name | Advanced biosensing and bioanalysis techniques |
| topics[2].id | https://openalex.org/T10725 |
| topics[2].field.id | https://openalex.org/fields/13 |
| topics[2].field.display_name | Biochemistry, Genetics and Molecular Biology |
| topics[2].score | 0.9832000136375427 |
| topics[2].domain.id | https://openalex.org/domains/1 |
| topics[2].domain.display_name | Life Sciences |
| topics[2].subfield.id | https://openalex.org/subfields/1312 |
| topics[2].subfield.display_name | Molecular Biology |
| topics[2].display_name | RNA Interference and Gene Delivery |
| funders[0].id | https://openalex.org/F4320321001 |
| funders[0].ror | https://ror.org/01h0zpd94 |
| funders[0].display_name | National Natural Science Foundation of China |
| funders[1].id | https://openalex.org/F4320321543 |
| funders[1].ror | https://ror.org/0426zh255 |
| funders[1].display_name | China Postdoctoral Science Foundation |
| funders[2].id | https://openalex.org/F4320335787 |
| funders[2].ror | |
| funders[2].display_name | Fundamental Research Funds for the Central Universities |
| funders[3].id | https://openalex.org/F4320336567 |
| funders[3].ror | |
| funders[3].display_name | Natural Science Basic Research Program of Shaanxi Province |
| is_xpac | False |
| apc_list.value | 980 |
| apc_list.currency | USD |
| apc_list.value_usd | 980 |
| apc_paid.value | 980 |
| apc_paid.currency | USD |
| apc_paid.value_usd | 980 |
| concepts[0].id | https://openalex.org/C22744801 |
| concepts[0].level | 3 |
| concepts[0].score | 0.8607355356216431 |
| concepts[0].wikidata | https://www.wikidata.org/wiki/Q172778 |
| concepts[0].display_name | Plasmid |
| concepts[1].id | https://openalex.org/C98108389 |
| concepts[1].level | 3 |
| concepts[1].score | 0.7702800631523132 |
| concepts[1].wikidata | https://www.wikidata.org/wiki/Q412563 |
| concepts[1].display_name | CRISPR |
| concepts[2].id | https://openalex.org/C16318435 |
| concepts[2].level | 4 |
| concepts[2].score | 0.7395537495613098 |
| concepts[2].wikidata | https://www.wikidata.org/wiki/Q149299 |
| concepts[2].display_name | Mutagenesis |
| concepts[3].id | https://openalex.org/C132455925 |
| concepts[3].level | 4 |
| concepts[3].score | 0.6178107857704163 |
| concepts[3].wikidata | https://www.wikidata.org/wiki/Q16965677 |
| concepts[3].display_name | Cas9 |
| concepts[4].id | https://openalex.org/C86803240 |
| concepts[4].level | 0 |
| concepts[4].score | 0.6166317462921143 |
| concepts[4].wikidata | https://www.wikidata.org/wiki/Q420 |
| concepts[4].display_name | Biology |
| concepts[5].id | https://openalex.org/C70721500 |
| concepts[5].level | 1 |
| concepts[5].score | 0.5027101039886475 |
| concepts[5].wikidata | https://www.wikidata.org/wiki/Q177005 |
| concepts[5].display_name | Computational biology |
| concepts[6].id | https://openalex.org/C54355233 |
| concepts[6].level | 1 |
| concepts[6].score | 0.4961443841457367 |
| concepts[6].wikidata | https://www.wikidata.org/wiki/Q7162 |
| concepts[6].display_name | Genetics |
| concepts[7].id | https://openalex.org/C144501496 |
| concepts[7].level | 4 |
| concepts[7].score | 0.48560088872909546 |
| concepts[7].wikidata | https://www.wikidata.org/wiki/Q5533489 |
| concepts[7].display_name | Genome editing |
| concepts[8].id | https://openalex.org/C163005120 |
| concepts[8].level | 4 |
| concepts[8].score | 0.4161298871040344 |
| concepts[8].wikidata | https://www.wikidata.org/wiki/Q4115252 |
| concepts[8].display_name | Subcloning |
| concepts[9].id | https://openalex.org/C552990157 |
| concepts[9].level | 2 |
| concepts[9].score | 0.3694046139717102 |
| concepts[9].wikidata | https://www.wikidata.org/wiki/Q7430 |
| concepts[9].display_name | DNA |
| concepts[10].id | https://openalex.org/C501734568 |
| concepts[10].level | 3 |
| concepts[10].score | 0.3061995804309845 |
| concepts[10].wikidata | https://www.wikidata.org/wiki/Q42918 |
| concepts[10].display_name | Mutation |
| concepts[11].id | https://openalex.org/C104317684 |
| concepts[11].level | 2 |
| concepts[11].score | 0.2765663266181946 |
| concepts[11].wikidata | https://www.wikidata.org/wiki/Q7187 |
| concepts[11].display_name | Gene |
| keywords[0].id | https://openalex.org/keywords/plasmid |
| keywords[0].score | 0.8607355356216431 |
| keywords[0].display_name | Plasmid |
| keywords[1].id | https://openalex.org/keywords/crispr |
| keywords[1].score | 0.7702800631523132 |
| keywords[1].display_name | CRISPR |
| keywords[2].id | https://openalex.org/keywords/mutagenesis |
| keywords[2].score | 0.7395537495613098 |
| keywords[2].display_name | Mutagenesis |
| keywords[3].id | https://openalex.org/keywords/cas9 |
| keywords[3].score | 0.6178107857704163 |
| keywords[3].display_name | Cas9 |
| keywords[4].id | https://openalex.org/keywords/biology |
| keywords[4].score | 0.6166317462921143 |
| keywords[4].display_name | Biology |
| keywords[5].id | https://openalex.org/keywords/computational-biology |
| keywords[5].score | 0.5027101039886475 |
| keywords[5].display_name | Computational biology |
| keywords[6].id | https://openalex.org/keywords/genetics |
| keywords[6].score | 0.4961443841457367 |
| keywords[6].display_name | Genetics |
| keywords[7].id | https://openalex.org/keywords/genome-editing |
| keywords[7].score | 0.48560088872909546 |
| keywords[7].display_name | Genome editing |
| keywords[8].id | https://openalex.org/keywords/subcloning |
| keywords[8].score | 0.4161298871040344 |
| keywords[8].display_name | Subcloning |
| keywords[9].id | https://openalex.org/keywords/dna |
| keywords[9].score | 0.3694046139717102 |
| keywords[9].display_name | DNA |
| keywords[10].id | https://openalex.org/keywords/mutation |
| keywords[10].score | 0.3061995804309845 |
| keywords[10].display_name | Mutation |
| keywords[11].id | https://openalex.org/keywords/gene |
| keywords[11].score | 0.2765663266181946 |
| keywords[11].display_name | Gene |
| language | en |
| locations[0].id | doi:10.1016/j.fmre.2022.06.017 |
| locations[0].is_oa | True |
| locations[0].source.id | https://openalex.org/S4210177083 |
| locations[0].source.issn | 2096-9457, 2667-3258 |
| locations[0].source.type | journal |
| locations[0].source.is_oa | True |
| locations[0].source.issn_l | 2096-9457 |
| locations[0].source.is_core | True |
| locations[0].source.is_in_doaj | True |
| locations[0].source.display_name | Fundamental Research |
| locations[0].source.host_organization | https://openalex.org/P4310320990 |
| locations[0].source.host_organization_name | Elsevier BV |
| locations[0].source.host_organization_lineage | https://openalex.org/P4310320990 |
| locations[0].source.host_organization_lineage_names | Elsevier BV |
| locations[0].license | cc-by-nc-nd |
| locations[0].pdf_url | |
| locations[0].version | publishedVersion |
| locations[0].raw_type | journal-article |
| locations[0].license_id | https://openalex.org/licenses/cc-by-nc-nd |
| locations[0].is_accepted | True |
| locations[0].is_published | True |
| locations[0].raw_source_name | Fundamental Research |
| locations[0].landing_page_url | https://doi.org/10.1016/j.fmre.2022.06.017 |
| locations[1].id | pmid:40777797 |
| locations[1].is_oa | False |
| locations[1].source.id | https://openalex.org/S4306525036 |
| locations[1].source.issn | |
| locations[1].source.type | repository |
| locations[1].source.is_oa | False |
| locations[1].source.issn_l | |
| locations[1].source.is_core | False |
| locations[1].source.is_in_doaj | False |
| locations[1].source.display_name | PubMed |
| locations[1].source.host_organization | https://openalex.org/I1299303238 |
| locations[1].source.host_organization_name | National Institutes of Health |
| locations[1].source.host_organization_lineage | https://openalex.org/I1299303238 |
| locations[1].license | |
| locations[1].pdf_url | |
| locations[1].version | publishedVersion |
| locations[1].raw_type | |
| locations[1].license_id | |
| locations[1].is_accepted | True |
| locations[1].is_published | True |
| locations[1].raw_source_name | Fundamental research |
| locations[1].landing_page_url | https://pubmed.ncbi.nlm.nih.gov/40777797 |
| locations[2].id | pmh:oai:doaj.org/article:90339c8870cf454aaa9deddcdb966782 |
| locations[2].is_oa | False |
| locations[2].source.id | https://openalex.org/S4306401280 |
| locations[2].source.issn | |
| locations[2].source.type | repository |
| locations[2].source.is_oa | False |
| locations[2].source.issn_l | |
| locations[2].source.is_core | False |
| locations[2].source.is_in_doaj | False |
| locations[2].source.display_name | DOAJ (DOAJ: Directory of Open Access Journals) |
| locations[2].source.host_organization | |
| locations[2].source.host_organization_name | |
| locations[2].license | |
| locations[2].pdf_url | |
| locations[2].version | submittedVersion |
| locations[2].raw_type | article |
| locations[2].license_id | |
| locations[2].is_accepted | False |
| locations[2].is_published | False |
| locations[2].raw_source_name | Fundamental Research, Vol 5, Iss 4, Pp 1481-1487 (2025) |
| locations[2].landing_page_url | https://doaj.org/article/90339c8870cf454aaa9deddcdb966782 |
| locations[3].id | pmh:oai:pubmedcentral.nih.gov:12327819 |
| locations[3].is_oa | True |
| locations[3].source.id | https://openalex.org/S2764455111 |
| locations[3].source.issn | |
| locations[3].source.type | repository |
| locations[3].source.is_oa | False |
| locations[3].source.issn_l | |
| locations[3].source.is_core | False |
| locations[3].source.is_in_doaj | False |
| locations[3].source.display_name | PubMed Central |
| locations[3].source.host_organization | https://openalex.org/I1299303238 |
| locations[3].source.host_organization_name | National Institutes of Health |
| locations[3].source.host_organization_lineage | https://openalex.org/I1299303238 |
| locations[3].license | other-oa |
| locations[3].pdf_url | |
| locations[3].version | submittedVersion |
| locations[3].raw_type | Text |
| locations[3].license_id | https://openalex.org/licenses/other-oa |
| locations[3].is_accepted | False |
| locations[3].is_published | False |
| locations[3].raw_source_name | Fundam Res |
| locations[3].landing_page_url | https://www.ncbi.nlm.nih.gov/pmc/articles/12327819 |
| indexed_in | crossref, doaj, pubmed |
| authorships[0].author.id | https://openalex.org/A5101434232 |
| authorships[0].author.orcid | https://orcid.org/0000-0001-8691-0889 |
| authorships[0].author.display_name | Fengjiao Zhao |
| authorships[0].countries | CN |
| authorships[0].affiliations[0].institution_ids | https://openalex.org/I87445476 |
| authorships[0].affiliations[0].raw_affiliation_string | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[0].institutions[0].id | https://openalex.org/I87445476 |
| authorships[0].institutions[0].ror | https://ror.org/017zhmm22 |
| authorships[0].institutions[0].type | education |
| authorships[0].institutions[0].lineage | https://openalex.org/I87445476 |
| authorships[0].institutions[0].country_code | CN |
| authorships[0].institutions[0].display_name | Xi'an Jiaotong University |
| authorships[0].author_position | first |
| authorships[0].raw_author_name | Fengjiao Zhao |
| authorships[0].is_corresponding | False |
| authorships[0].raw_affiliation_strings | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[1].author.id | https://openalex.org/A5100640421 |
| authorships[1].author.orcid | https://orcid.org/0000-0003-0188-6256 |
| authorships[1].author.display_name | Feng Chen |
| authorships[1].countries | CN |
| authorships[1].affiliations[0].institution_ids | https://openalex.org/I87445476 |
| authorships[1].affiliations[0].raw_affiliation_string | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[1].institutions[0].id | https://openalex.org/I87445476 |
| authorships[1].institutions[0].ror | https://ror.org/017zhmm22 |
| authorships[1].institutions[0].type | education |
| authorships[1].institutions[0].lineage | https://openalex.org/I87445476 |
| authorships[1].institutions[0].country_code | CN |
| authorships[1].institutions[0].display_name | Xi'an Jiaotong University |
| authorships[1].author_position | middle |
| authorships[1].raw_author_name | Feng Chen |
| authorships[1].is_corresponding | False |
| authorships[1].raw_affiliation_strings | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[2].author.id | https://openalex.org/A5055193360 |
| authorships[2].author.orcid | |
| authorships[2].author.display_name | Huahang Yu |
| authorships[2].countries | CN |
| authorships[2].affiliations[0].institution_ids | https://openalex.org/I87445476 |
| authorships[2].affiliations[0].raw_affiliation_string | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[2].institutions[0].id | https://openalex.org/I87445476 |
| authorships[2].institutions[0].ror | https://ror.org/017zhmm22 |
| authorships[2].institutions[0].type | education |
| authorships[2].institutions[0].lineage | https://openalex.org/I87445476 |
| authorships[2].institutions[0].country_code | CN |
| authorships[2].institutions[0].display_name | Xi'an Jiaotong University |
| authorships[2].author_position | middle |
| authorships[2].raw_author_name | Huahang Yu |
| authorships[2].is_corresponding | False |
| authorships[2].raw_affiliation_strings | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[3].author.id | https://openalex.org/A5050924152 |
| authorships[3].author.orcid | https://orcid.org/0000-0001-7916-9332 |
| authorships[3].author.display_name | Siyue Fan |
| authorships[3].countries | CN |
| authorships[3].affiliations[0].institution_ids | https://openalex.org/I87445476 |
| authorships[3].affiliations[0].raw_affiliation_string | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[3].institutions[0].id | https://openalex.org/I87445476 |
| authorships[3].institutions[0].ror | https://ror.org/017zhmm22 |
| authorships[3].institutions[0].type | education |
| authorships[3].institutions[0].lineage | https://openalex.org/I87445476 |
| authorships[3].institutions[0].country_code | CN |
| authorships[3].institutions[0].display_name | Xi'an Jiaotong University |
| authorships[3].author_position | middle |
| authorships[3].raw_author_name | Siyue Fan |
| authorships[3].is_corresponding | False |
| authorships[3].raw_affiliation_strings | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[4].author.id | https://openalex.org/A5101003336 |
| authorships[4].author.orcid | |
| authorships[4].author.display_name | Bai Min |
| authorships[4].countries | CN |
| authorships[4].affiliations[0].institution_ids | https://openalex.org/I87445476 |
| authorships[4].affiliations[0].raw_affiliation_string | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[4].institutions[0].id | https://openalex.org/I87445476 |
| authorships[4].institutions[0].ror | https://ror.org/017zhmm22 |
| authorships[4].institutions[0].type | education |
| authorships[4].institutions[0].lineage | https://openalex.org/I87445476 |
| authorships[4].institutions[0].country_code | CN |
| authorships[4].institutions[0].display_name | Xi'an Jiaotong University |
| authorships[4].author_position | middle |
| authorships[4].raw_author_name | Min Bai |
| authorships[4].is_corresponding | False |
| authorships[4].raw_affiliation_strings | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[5].author.id | https://openalex.org/A5100763210 |
| authorships[5].author.orcid | https://orcid.org/0000-0001-9259-4325 |
| authorships[5].author.display_name | Jing Xue |
| authorships[5].countries | CN |
| authorships[5].affiliations[0].institution_ids | https://openalex.org/I87445476 |
| authorships[5].affiliations[0].raw_affiliation_string | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[5].institutions[0].id | https://openalex.org/I87445476 |
| authorships[5].institutions[0].ror | https://ror.org/017zhmm22 |
| authorships[5].institutions[0].type | education |
| authorships[5].institutions[0].lineage | https://openalex.org/I87445476 |
| authorships[5].institutions[0].country_code | CN |
| authorships[5].institutions[0].display_name | Xi'an Jiaotong University |
| authorships[5].author_position | middle |
| authorships[5].raw_author_name | Jing Xue |
| authorships[5].is_corresponding | False |
| authorships[5].raw_affiliation_strings | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[6].author.id | https://openalex.org/A5081257424 |
| authorships[6].author.orcid | https://orcid.org/0000-0001-5544-5697 |
| authorships[6].author.display_name | Yue Zhao |
| authorships[6].countries | CN |
| authorships[6].affiliations[0].institution_ids | https://openalex.org/I87445476 |
| authorships[6].affiliations[0].raw_affiliation_string | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[6].institutions[0].id | https://openalex.org/I87445476 |
| authorships[6].institutions[0].ror | https://ror.org/017zhmm22 |
| authorships[6].institutions[0].type | education |
| authorships[6].institutions[0].lineage | https://openalex.org/I87445476 |
| authorships[6].institutions[0].country_code | CN |
| authorships[6].institutions[0].display_name | Xi'an Jiaotong University |
| authorships[6].author_position | middle |
| authorships[6].raw_author_name | Yue Zhao |
| authorships[6].is_corresponding | False |
| authorships[6].raw_affiliation_strings | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[7].author.id | https://openalex.org/A5047923935 |
| authorships[7].author.orcid | https://orcid.org/0000-0001-7505-2727 |
| authorships[7].author.display_name | Xiaolei Zuo |
| authorships[7].countries | CN |
| authorships[7].affiliations[0].institution_ids | https://openalex.org/I183067930 |
| authorships[7].affiliations[0].raw_affiliation_string | School of Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules and National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai 200240, China |
| authorships[7].institutions[0].id | https://openalex.org/I183067930 |
| authorships[7].institutions[0].ror | https://ror.org/0220qvk04 |
| authorships[7].institutions[0].type | education |
| authorships[7].institutions[0].lineage | https://openalex.org/I183067930 |
| authorships[7].institutions[0].country_code | CN |
| authorships[7].institutions[0].display_name | Shanghai Jiao Tong University |
| authorships[7].author_position | middle |
| authorships[7].raw_author_name | Xiaolei Zuo |
| authorships[7].is_corresponding | False |
| authorships[7].raw_affiliation_strings | School of Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules and National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai 200240, China |
| authorships[8].author.id | https://openalex.org/A5006228310 |
| authorships[8].author.orcid | https://orcid.org/0000-0002-7171-7338 |
| authorships[8].author.display_name | Chunhai Fan |
| authorships[8].countries | CN |
| authorships[8].affiliations[0].institution_ids | https://openalex.org/I183067930 |
| authorships[8].affiliations[0].raw_affiliation_string | School of Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules and National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai 200240, China |
| authorships[8].institutions[0].id | https://openalex.org/I183067930 |
| authorships[8].institutions[0].ror | https://ror.org/0220qvk04 |
| authorships[8].institutions[0].type | education |
| authorships[8].institutions[0].lineage | https://openalex.org/I183067930 |
| authorships[8].institutions[0].country_code | CN |
| authorships[8].institutions[0].display_name | Shanghai Jiao Tong University |
| authorships[8].author_position | middle |
| authorships[8].raw_author_name | Chunhai Fan |
| authorships[8].is_corresponding | False |
| authorships[8].raw_affiliation_strings | School of Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules and National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai 200240, China |
| authorships[9].author.id | https://openalex.org/A5057672960 |
| authorships[9].author.orcid | https://orcid.org/0000-0002-1796-7651 |
| authorships[9].author.display_name | Yongxi Zhao |
| authorships[9].countries | CN |
| authorships[9].affiliations[0].institution_ids | https://openalex.org/I87445476 |
| authorships[9].affiliations[0].raw_affiliation_string | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| authorships[9].institutions[0].id | https://openalex.org/I87445476 |
| authorships[9].institutions[0].ror | https://ror.org/017zhmm22 |
| authorships[9].institutions[0].type | education |
| authorships[9].institutions[0].lineage | https://openalex.org/I87445476 |
| authorships[9].institutions[0].country_code | CN |
| authorships[9].institutions[0].display_name | Xi'an Jiaotong University |
| authorships[9].author_position | last |
| authorships[9].raw_author_name | Yongxi Zhao |
| authorships[9].is_corresponding | True |
| authorships[9].raw_affiliation_strings | Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, Shaanxi 710049, China |
| has_content.pdf | False |
| has_content.grobid_xml | False |
| is_paratext | False |
| open_access.is_oa | True |
| open_access.oa_url | https://doi.org/10.1016/j.fmre.2022.06.017 |
| open_access.oa_status | gold |
| open_access.any_repository_has_fulltext | False |
| created_date | 2025-10-10T00:00:00 |
| display_name | CRISPR/Cas system-guided plasmid mutagenesis without sequence restriction |
| has_fulltext | False |
| is_retracted | False |
| updated_date | 2025-11-06T03:46:38.306776 |
| primary_topic.id | https://openalex.org/T10878 |
| primary_topic.field.id | https://openalex.org/fields/13 |
| primary_topic.field.display_name | Biochemistry, Genetics and Molecular Biology |
| primary_topic.score | 1.0 |
| primary_topic.domain.id | https://openalex.org/domains/1 |
| primary_topic.domain.display_name | Life Sciences |
| primary_topic.subfield.id | https://openalex.org/subfields/1312 |
| primary_topic.subfield.display_name | Molecular Biology |
| primary_topic.display_name | CRISPR and Genetic Engineering |
| related_works | https://openalex.org/W4366822718, https://openalex.org/W4311501380, https://openalex.org/W2603161682, https://openalex.org/W2807464733, https://openalex.org/W2516842346, https://openalex.org/W2903635918, https://openalex.org/W2437118042, https://openalex.org/W3052741125, https://openalex.org/W2466284841, https://openalex.org/W2357872089 |
| cited_by_count | 0 |
| locations_count | 4 |
| best_oa_location.id | doi:10.1016/j.fmre.2022.06.017 |
| best_oa_location.is_oa | True |
| best_oa_location.source.id | https://openalex.org/S4210177083 |
| best_oa_location.source.issn | 2096-9457, 2667-3258 |
| best_oa_location.source.type | journal |
| best_oa_location.source.is_oa | True |
| best_oa_location.source.issn_l | 2096-9457 |
| best_oa_location.source.is_core | True |
| best_oa_location.source.is_in_doaj | True |
| best_oa_location.source.display_name | Fundamental Research |
| best_oa_location.source.host_organization | https://openalex.org/P4310320990 |
| best_oa_location.source.host_organization_name | Elsevier BV |
| best_oa_location.source.host_organization_lineage | https://openalex.org/P4310320990 |
| best_oa_location.source.host_organization_lineage_names | Elsevier BV |
| best_oa_location.license | cc-by-nc-nd |
| best_oa_location.pdf_url | |
| best_oa_location.version | publishedVersion |
| best_oa_location.raw_type | journal-article |
| best_oa_location.license_id | https://openalex.org/licenses/cc-by-nc-nd |
| best_oa_location.is_accepted | True |
| best_oa_location.is_published | True |
| best_oa_location.raw_source_name | Fundamental Research |
| best_oa_location.landing_page_url | https://doi.org/10.1016/j.fmre.2022.06.017 |
| primary_location.id | doi:10.1016/j.fmre.2022.06.017 |
| primary_location.is_oa | True |
| primary_location.source.id | https://openalex.org/S4210177083 |
| primary_location.source.issn | 2096-9457, 2667-3258 |
| primary_location.source.type | journal |
| primary_location.source.is_oa | True |
| primary_location.source.issn_l | 2096-9457 |
| primary_location.source.is_core | True |
| primary_location.source.is_in_doaj | True |
| primary_location.source.display_name | Fundamental Research |
| primary_location.source.host_organization | https://openalex.org/P4310320990 |
| primary_location.source.host_organization_name | Elsevier BV |
| primary_location.source.host_organization_lineage | https://openalex.org/P4310320990 |
| primary_location.source.host_organization_lineage_names | Elsevier BV |
| primary_location.license | cc-by-nc-nd |
| primary_location.pdf_url | |
| primary_location.version | publishedVersion |
| primary_location.raw_type | journal-article |
| primary_location.license_id | https://openalex.org/licenses/cc-by-nc-nd |
| primary_location.is_accepted | True |
| primary_location.is_published | True |
| primary_location.raw_source_name | Fundamental Research |
| primary_location.landing_page_url | https://doi.org/10.1016/j.fmre.2022.06.017 |
| publication_date | 2022-07-15 |
| publication_year | 2022 |
| referenced_works | https://openalex.org/W2913425582, https://openalex.org/W3138025811, https://openalex.org/W2810494969, https://openalex.org/W2118265845, https://openalex.org/W2529766297, https://openalex.org/W2961875637, https://openalex.org/W4220822979, https://openalex.org/W1552716706, https://openalex.org/W2160253662, https://openalex.org/W1971439989, https://openalex.org/W2133582783, https://openalex.org/W2084615238, https://openalex.org/W4210539126, https://openalex.org/W1977463604, https://openalex.org/W2013583230, https://openalex.org/W2077892824, https://openalex.org/W2606606197, https://openalex.org/W3035141594, https://openalex.org/W2042655455, https://openalex.org/W1988156863, https://openalex.org/W2007847817, https://openalex.org/W2168103157, https://openalex.org/W2530338180, https://openalex.org/W2771499133, https://openalex.org/W2947777875, https://openalex.org/W2888908334, https://openalex.org/W2770127492, https://openalex.org/W1946097599, https://openalex.org/W2788069741, https://openalex.org/W3108602813, https://openalex.org/W2916530321, https://openalex.org/W3111023718, https://openalex.org/W6601859127, https://openalex.org/W1964111587, https://openalex.org/W2903602386, https://openalex.org/W2165869860, https://openalex.org/W3201162411, https://openalex.org/W2788728311, https://openalex.org/W2889680755, https://openalex.org/W2622596320, https://openalex.org/W2302008724, https://openalex.org/W2926667013, https://openalex.org/W2620475836, https://openalex.org/W3013504322, https://openalex.org/W2950281100, https://openalex.org/W4245041385 |
| referenced_works_count | 46 |
| abstract_inverted_index.a | 20, 51 |
| abstract_inverted_index.It | 78 |
| abstract_inverted_index.We | 88 |
| abstract_inverted_index.an | 3, 138 |
| abstract_inverted_index.as | 62 |
| abstract_inverted_index.in | 36, 106 |
| abstract_inverted_index.is | 2, 19 |
| abstract_inverted_index.it | 30 |
| abstract_inverted_index.of | 15, 39 |
| abstract_inverted_index.on | 17, 75 |
| abstract_inverted_index.to | 6, 24, 57, 96 |
| abstract_inverted_index.we | 44 |
| abstract_inverted_index.PAM | 116, 126 |
| abstract_inverted_index.The | 101 |
| abstract_inverted_index.and | 27, 83, 118, 128, 144 |
| abstract_inverted_index.can | 69, 124 |
| abstract_inverted_index.for | 65, 132, 141 |
| abstract_inverted_index.the | 33, 37, 46, 76, 107, 115 |
| abstract_inverted_index.yet | 29 |
| abstract_inverted_index.Here | 43 |
| abstract_inverted_index.This | 67, 135 |
| abstract_inverted_index.even | 84 |
| abstract_inverted_index.from | 32 |
| abstract_inverted_index.step | 5 |
| abstract_inverted_index.this | 91 |
| abstract_inverted_index.tool | 140 |
| abstract_inverted_index.were | 110 |
| abstract_inverted_index.with | 10, 93, 104 |
| abstract_inverted_index.work | 136 |
| abstract_inverted_index.broad | 80 |
| abstract_inverted_index.lower | 120 |
| abstract_inverted_index.range | 117 |
| abstract_inverted_index.using | 53 |
| abstract_inverted_index.which | 123 |
| abstract_inverted_index.Direct | 13 |
| abstract_inverted_index.FnCpf1 | 102 |
| abstract_inverted_index.covers | 85 |
| abstract_inverted_index.create | 71 |
| abstract_inverted_index.design | 95 |
| abstract_inverted_index.genome | 133 |
| abstract_inverted_index.groove | 109 |
| abstract_inverted_index.loosen | 125 |
| abstract_inverted_index.method | 22, 52, 68, 92 |
| abstract_inverted_index.offers | 79 |
| abstract_inverted_index.target | 130 |
| abstract_inverted_index.FnCpf1. | 100 |
| abstract_inverted_index.Plasmid | 0 |
| abstract_inverted_index.Several | 112 |
| abstract_inverted_index.combine | 90 |
| abstract_inverted_index.complex | 56 |
| abstract_inverted_index.desired | 11 |
| abstract_inverted_index.effect, | 122 |
| abstract_inverted_index.enhance | 129 |
| abstract_inverted_index.further | 89 |
| abstract_inverted_index.methods | 26 |
| abstract_inverted_index.nickase | 55 |
| abstract_inverted_index.plasmid | 49, 142 |
| abstract_inverted_index.protein | 8, 99, 145 |
| abstract_inverted_index.suffers | 31 |
| abstract_inverted_index.circular | 41, 60 |
| abstract_inverted_index.compared | 23 |
| abstract_inverted_index.directly | 70 |
| abstract_inverted_index.editing. | 134 |
| abstract_inverted_index.engineer | 7, 97 |
| abstract_inverted_index.expanded | 114 |
| abstract_inverted_index.generate | 58 |
| abstract_inverted_index.mutation | 73 |
| abstract_inverted_index.plasmid. | 77 |
| abstract_inverted_index.plasmids | 18, 61 |
| abstract_inverted_index.presents | 137 |
| abstract_inverted_index.rational | 94 |
| abstract_inverted_index.sequence | 34, 81 |
| abstract_inverted_index.studies. | 147 |
| abstract_inverted_index.variants | 9, 103, 113 |
| abstract_inverted_index.PCR-based | 25 |
| abstract_inverted_index.effective | 139 |
| abstract_inverted_index.essential | 4 |
| abstract_inverted_index.exhibited | 119 |
| abstract_inverted_index.gRNA/Cas9 | 54 |
| abstract_inverted_index.libraries | 74 |
| abstract_inverted_index.mutations | 16, 105 |
| abstract_inverted_index.plasmids. | 42, 87 |
| abstract_inverted_index.promising | 21 |
| abstract_inverted_index.templates | 64 |
| abstract_inverted_index.CRISPR/Cas | 47 |
| abstract_inverted_index.constraint | 127 |
| abstract_inverted_index.generated. | 111 |
| abstract_inverted_index.generation | 14, 38 |
| abstract_inverted_index.methylated | 86 |
| abstract_inverted_index.off-target | 121 |
| abstract_inverted_index.PAM-binding | 108 |
| abstract_inverted_index.demonstrate | 45 |
| abstract_inverted_index.engineering | 146 |
| abstract_inverted_index.mutagenesis | 1, 143 |
| abstract_inverted_index.properties. | 12 |
| abstract_inverted_index.restriction | 35 |
| abstract_inverted_index.specificity | 131 |
| abstract_inverted_index.subcloning, | 28 |
| abstract_inverted_index.mutagenesis, | 50 |
| abstract_inverted_index.mutagenesis. | 66 |
| abstract_inverted_index.user-defined | 72 |
| abstract_inverted_index.system-guided | 48 |
| abstract_inverted_index.genome-editing | 98 |
| abstract_inverted_index.polymerization | 63 |
| abstract_inverted_index.programmability | 82 |
| abstract_inverted_index.single-stranded | 40, 59 |
| cited_by_percentile_year | |
| corresponding_author_ids | https://openalex.org/A5057672960 |
| countries_distinct_count | 1 |
| institutions_distinct_count | 10 |
| corresponding_institution_ids | https://openalex.org/I87445476 |
| citation_normalized_percentile.value | 0.08292347 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |