CRISPR-Cas9 recognition of enzymatically synthesized base-modified nucleic acids Article Swipe
Hui Yang
,
Elena Eremeeva
,
Mikhail Abramov
,
Maarten Jacquemyn
,
Elisabetta Groaz
,
Dirk Daelemans
,
Piet Herdewijn
·
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1093/nar/gkac1147
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1093/nar/gkac1147
An enzymatic method has been successfully established enabling the generation of partially base-modified RNA (previously named RZA) constructs, in which all G residues were replaced by isomorphic fluorescent thienoguanosine (thG) analogs, as well as fully modified RZA featuring thG, 5-bromocytosine, 7-deazaadenine and 5-chlorouracil. The transcriptional efficiency of emissive fully modified RZA was found to benefit from the use of various T7 RNA polymerase variants. Moreover, dthG could be incorporated into PCR products by Taq DNA polymerase together with the other three base-modified nucleotides. Notably, the obtained RNA products containing thG as well as thG together with 5-bromocytosine could function as effectively as natural sgRNAs in an in vitro CRISPR-Cas9 cleavage assay. N1-Methylpseudouridine was also demonstrated to be a faithful non-canonical substitute of uridine to direct Cas9 nuclease cleavage when incorporated in sgRNA. The Cas9 inactivation by 7-deazapurines indicated the importance of the 7-nitrogen atom of purines in both sgRNA and PAM site for achieving efficient Cas9 cleavage. Additional aspects of this study are discussed in relation to the significance of sgRNA–protein and PAM–-protein interactions that were not highlighted by the Cas9–sgRNA–DNA complex crystal structure. These findings could expand the impact and therapeutic value of CRISPR-Cas9 and other RNA-based technologies.
Related Topics
Concepts
Cas9
Guide RNA
CRISPR
Biology
Subgenomic mRNA
Genome editing
RNA
Nuclease
DNA
Nucleic acid
Gene
Molecular biology
T7 RNA polymerase
Computational biology
Biochemistry
Escherichia coli
Bacteriophage
Metadata
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- article
- Language
- en
- Landing Page
- https://doi.org/10.1093/nar/gkac1147
- https://academic.oup.com/nar/article-pdf/51/4/1501/50423672/gkac1147.pdf
- OA Status
- gold
- Cited By
- 15
- References
- 52
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4313706607
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4313706607Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1093/nar/gkac1147Digital Object Identifier
- Title
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CRISPR-Cas9 recognition of enzymatically synthesized base-modified nucleic acidsWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
-
2023Year of publication
- Publication date
-
2023-01-06Full publication date if available
- Authors
-
Hui Yang, Elena Eremeeva, Mikhail Abramov, Maarten Jacquemyn, Elisabetta Groaz, Dirk Daelemans, Piet HerdewijnList of authors in order
- Landing page
-
https://doi.org/10.1093/nar/gkac1147Publisher landing page
- PDF URL
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https://academic.oup.com/nar/article-pdf/51/4/1501/50423672/gkac1147.pdfDirect link to full text PDF
- Open access
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YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
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https://academic.oup.com/nar/article-pdf/51/4/1501/50423672/gkac1147.pdfDirect OA link when available
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Cas9, Guide RNA, CRISPR, Biology, Subgenomic mRNA, Genome editing, RNA, Nuclease, DNA, Nucleic acid, Gene, Molecular biology, T7 RNA polymerase, Computational biology, Biochemistry, Escherichia coli, BacteriophageTop concepts (fields/topics) attached by OpenAlex
- Cited by
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15Total citation count in OpenAlex
- Citations by year (recent)
-
2025: 4, 2024: 7, 2023: 4Per-year citation counts (last 5 years)
- References (count)
-
52Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.sgRNA | 149 |
| abstract_inverted_index.study | 162 |
| abstract_inverted_index.three | 81 |
| abstract_inverted_index.value | 193 |
| abstract_inverted_index.vitro | 108 |
| abstract_inverted_index.which | 20 |
| abstract_inverted_index.assay. | 111 |
| abstract_inverted_index.direct | 125 |
| abstract_inverted_index.expand | 188 |
| abstract_inverted_index.impact | 190 |
| abstract_inverted_index.method | 3 |
| abstract_inverted_index.sgRNA. | 132 |
| abstract_inverted_index.sgRNAs | 104 |
| abstract_inverted_index.aspects | 159 |
| abstract_inverted_index.benefit | 55 |
| abstract_inverted_index.complex | 182 |
| abstract_inverted_index.crystal | 183 |
| abstract_inverted_index.natural | 103 |
| abstract_inverted_index.purines | 146 |
| abstract_inverted_index.uridine | 123 |
| abstract_inverted_index.various | 60 |
| abstract_inverted_index.Abstract | 0 |
| abstract_inverted_index.Notably, | 84 |
| abstract_inverted_index.analogs, | 31 |
| abstract_inverted_index.cleavage | 110, 128 |
| abstract_inverted_index.emissive | 48 |
| abstract_inverted_index.enabling | 8 |
| abstract_inverted_index.faithful | 119 |
| abstract_inverted_index.findings | 186 |
| abstract_inverted_index.function | 99 |
| abstract_inverted_index.modified | 36, 50 |
| abstract_inverted_index.nuclease | 127 |
| abstract_inverted_index.obtained | 86 |
| abstract_inverted_index.products | 72, 88 |
| abstract_inverted_index.relation | 166 |
| abstract_inverted_index.replaced | 25 |
| abstract_inverted_index.residues | 23 |
| abstract_inverted_index.together | 77, 95 |
| abstract_inverted_index.Moreover, | 65 |
| abstract_inverted_index.RNA-based | 198 |
| abstract_inverted_index.achieving | 154 |
| abstract_inverted_index.cleavage. | 157 |
| abstract_inverted_index.discussed | 164 |
| abstract_inverted_index.efficient | 155 |
| abstract_inverted_index.enzymatic | 2 |
| abstract_inverted_index.featuring | 38 |
| abstract_inverted_index.indicated | 138 |
| abstract_inverted_index.partially | 12 |
| abstract_inverted_index.variants. | 64 |
| abstract_inverted_index.7-nitrogen | 143 |
| abstract_inverted_index.Additional | 158 |
| abstract_inverted_index.containing | 89 |
| abstract_inverted_index.efficiency | 46 |
| abstract_inverted_index.generation | 10 |
| abstract_inverted_index.importance | 140 |
| abstract_inverted_index.isomorphic | 27 |
| abstract_inverted_index.polymerase | 63, 76 |
| abstract_inverted_index.structure. | 184 |
| abstract_inverted_index.substitute | 121 |
| abstract_inverted_index.(previously | 15 |
| abstract_inverted_index.CRISPR-Cas9 | 109, 195 |
| abstract_inverted_index.constructs, | 18 |
| abstract_inverted_index.effectively | 101 |
| abstract_inverted_index.established | 7 |
| abstract_inverted_index.fluorescent | 28 |
| abstract_inverted_index.highlighted | 178 |
| abstract_inverted_index.therapeutic | 192 |
| abstract_inverted_index.demonstrated | 115 |
| abstract_inverted_index.inactivation | 135 |
| abstract_inverted_index.incorporated | 69, 130 |
| abstract_inverted_index.interactions | 174 |
| abstract_inverted_index.nucleotides. | 83 |
| abstract_inverted_index.significance | 169 |
| abstract_inverted_index.successfully | 6 |
| abstract_inverted_index.base-modified | 13, 82 |
| abstract_inverted_index.non-canonical | 120 |
| abstract_inverted_index.technologies. | 199 |
| abstract_inverted_index.7-deazaadenine | 41 |
| abstract_inverted_index.7-deazapurines | 137 |
| abstract_inverted_index.PAM–-protein | 173 |
| abstract_inverted_index.5-bromocytosine | 97 |
| abstract_inverted_index.5-chlorouracil. | 43 |
| abstract_inverted_index.sgRNA–protein | 171 |
| abstract_inverted_index.thienoguanosine | 29 |
| abstract_inverted_index.transcriptional | 45 |
| abstract_inverted_index.5-bromocytosine, | 40 |
| abstract_inverted_index.Cas9–sgRNA–DNA | 181 |
| abstract_inverted_index.N1-Methylpseudouridine | 112 |
| cited_by_percentile_year.max | 99 |
| cited_by_percentile_year.min | 97 |
| corresponding_author_ids | https://openalex.org/A5006255227 |
| countries_distinct_count | 3 |
| institutions_distinct_count | 7 |
| corresponding_institution_ids | https://openalex.org/I24156275, https://openalex.org/I99464096 |
| citation_normalized_percentile.value | 0.9013853 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | True |