Expanding the CRISPR Toolbox with ErCas12a in Zebrafish and Human Cells Article Swipe
Wesley A. Wierson
,
Brandon W. Simone
,
Zachary WareJoncas
,
Carla M. Mann
,
Jordan M. Welker
,
Bibekananda Kar
,
Michael J. Emch
,
Iddo Friedberg
,
William A.C. Gendron
,
Michael A. Barry
,
Karl J. Clark
,
Drena Dobbs
,
Maura McGrail
,
Stephen C. Ekker
,
Jeffrey J. Essner
·
YOU?
·
· 2019
· Open Access
·
· DOI: https://doi.org/10.1089/crispr.2019.0026
YOU?
·
· 2019
· Open Access
·
· DOI: https://doi.org/10.1089/crispr.2019.0026
CRISPR and CRISPR-Cas effector proteins enable the targeting of DNA double-strand breaks to defined loci based on a variable length RNA guide specific to each effector. The guide RNAs are generally similar in size and form, consisting of a ∼20 nucleotide sequence complementary to the DNA target and an RNA secondary structure recognized by the effector. However, the effector proteins vary in protospacer adjacent motif requirements, nuclease activities, and DNA binding kinetics. Recently, ErCas12a, a new member of the Cas12a family, was identified in Eubacterium rectale. Here, we report the first characterization of ErCas12a activity in zebrafish and expand on previously reported activity in human cells. Using a fluorescent reporter system, we show that CRISPR-ErCas12a elicits strand annealing mediated DNA repair more efficiently than CRISPR-Cas9. Further, using our previously reported gene targeting method that utilizes short homology, GeneWeld, we demonstrate the use of CRISPR-ErCas12a to integrate reporter alleles into the genomes of both zebrafish and human cells. Together, this work provides methods for deploying an additional CRISPR-Cas system, thus increasing the flexibility researchers have in applying genome engineering technologies.
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1089/crispr.2019.0026
- https://www.liebertpub.com/doi/pdf/10.1089/crispr.2019.0026
- OA Status
- hybrid
- Cited By
- 56
- References
- 58
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W2990128054
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
-
https://openalex.org/W2990128054Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.1089/crispr.2019.0026Digital Object Identifier
- Title
-
Expanding the CRISPR Toolbox with ErCas12a in Zebrafish and Human CellsWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2019Year of publication
- Publication date
-
2019-12-01Full publication date if available
- Authors
-
Wesley A. Wierson, Brandon W. Simone, Zachary WareJoncas, Carla M. Mann, Jordan M. Welker, Bibekananda Kar, Michael J. Emch, Iddo Friedberg, William A.C. Gendron, Michael A. Barry, Karl J. Clark, Drena Dobbs, Maura McGrail, Stephen C. Ekker, Jeffrey J. EssnerList of authors in order
- Landing page
-
https://doi.org/10.1089/crispr.2019.0026Publisher landing page
- PDF URL
-
https://www.liebertpub.com/doi/pdf/10.1089/crispr.2019.0026Direct link to full text PDF
- Open access
-
YesWhether a free full text is available
- OA status
-
hybridOpen access status per OpenAlex
- OA URL
-
https://www.liebertpub.com/doi/pdf/10.1089/crispr.2019.0026Direct OA link when available
- Concepts
-
CRISPR, Effector, Biology, Cas9, Nuclease, Genome editing, DNA, Computational biology, Genome engineering, RNA, Zebrafish, Gene, Guide RNA, Genome, Genetics, Cell biologyTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
56Total citation count in OpenAlex
- Citations by year (recent)
-
2025: 8, 2024: 6, 2023: 11, 2022: 11, 2021: 13Per-year citation counts (last 5 years)
- References (count)
-
58Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
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| counts_by_year[2].year | 2023 |
| counts_by_year[2].cited_by_count | 11 |
| counts_by_year[3].year | 2022 |
| counts_by_year[3].cited_by_count | 11 |
| counts_by_year[4].year | 2021 |
| counts_by_year[4].cited_by_count | 13 |
| counts_by_year[5].year | 2020 |
| counts_by_year[5].cited_by_count | 7 |
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| best_oa_location.source.host_organization_lineage | https://openalex.org/P4310320443 |
| best_oa_location.source.host_organization_lineage_names | Mary Ann Liebert, Inc. |
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| best_oa_location.is_published | True |
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| primary_location.source.is_oa | False |
| primary_location.source.issn_l | 2573-1599 |
| primary_location.source.is_core | True |
| primary_location.source.is_in_doaj | False |
| primary_location.source.display_name | The CRISPR Journal |
| primary_location.source.host_organization | https://openalex.org/P4310320443 |
| primary_location.source.host_organization_name | Mary Ann Liebert, Inc. |
| primary_location.source.host_organization_lineage | https://openalex.org/P4310320443 |
| primary_location.source.host_organization_lineage_names | Mary Ann Liebert, Inc. |
| primary_location.license | cc-by |
| primary_location.pdf_url | https://www.liebertpub.com/doi/pdf/10.1089/crispr.2019.0026 |
| primary_location.version | publishedVersion |
| primary_location.raw_type | journal-article |
| primary_location.license_id | https://openalex.org/licenses/cc-by |
| primary_location.is_accepted | True |
| primary_location.is_published | True |
| primary_location.raw_source_name | The CRISPR Journal |
| primary_location.landing_page_url | https://doi.org/10.1089/crispr.2019.0026 |
| publication_date | 2019-12-01 |
| publication_year | 2019 |
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| referenced_works_count | 58 |
| abstract_inverted_index.a | 17, 38, 74, 107 |
| abstract_inverted_index.an | 48, 164 |
| abstract_inverted_index.by | 53 |
| abstract_inverted_index.in | 32, 61, 83, 95, 103, 174 |
| abstract_inverted_index.of | 8, 37, 77, 92, 142, 151 |
| abstract_inverted_index.on | 16, 99 |
| abstract_inverted_index.to | 12, 23, 43, 144 |
| abstract_inverted_index.we | 87, 111, 138 |
| abstract_inverted_index.DNA | 9, 45, 69, 119 |
| abstract_inverted_index.RNA | 20, 49 |
| abstract_inverted_index.The | 26 |
| abstract_inverted_index.and | 1, 34, 47, 68, 97, 154 |
| abstract_inverted_index.are | 29 |
| abstract_inverted_index.for | 162 |
| abstract_inverted_index.new | 75 |
| abstract_inverted_index.our | 127 |
| abstract_inverted_index.the | 6, 44, 54, 57, 78, 89, 140, 149, 170 |
| abstract_inverted_index.use | 141 |
| abstract_inverted_index.was | 81 |
| abstract_inverted_index.RNAs | 28 |
| abstract_inverted_index.both | 152 |
| abstract_inverted_index.each | 24 |
| abstract_inverted_index.gene | 130 |
| abstract_inverted_index.have | 173 |
| abstract_inverted_index.into | 148 |
| abstract_inverted_index.loci | 14 |
| abstract_inverted_index.more | 121 |
| abstract_inverted_index.show | 112 |
| abstract_inverted_index.size | 33 |
| abstract_inverted_index.than | 123 |
| abstract_inverted_index.that | 113, 133 |
| abstract_inverted_index.this | 158 |
| abstract_inverted_index.thus | 168 |
| abstract_inverted_index.vary | 60 |
| abstract_inverted_index.work | 159 |
| abstract_inverted_index.Here, | 86 |
| abstract_inverted_index.Using | 106 |
| abstract_inverted_index.based | 15 |
| abstract_inverted_index.first | 90 |
| abstract_inverted_index.form, | 35 |
| abstract_inverted_index.guide | 21, 27 |
| abstract_inverted_index.human | 104, 155 |
| abstract_inverted_index.motif | 64 |
| abstract_inverted_index.short | 135 |
| abstract_inverted_index.using | 126 |
| abstract_inverted_index.∼20 | 39 |
| abstract_inverted_index.CRISPR | 0 |
| abstract_inverted_index.Cas12a | 79 |
| abstract_inverted_index.breaks | 11 |
| abstract_inverted_index.cells. | 105, 156 |
| abstract_inverted_index.enable | 5 |
| abstract_inverted_index.expand | 98 |
| abstract_inverted_index.genome | 176 |
| abstract_inverted_index.length | 19 |
| abstract_inverted_index.member | 76 |
| abstract_inverted_index.method | 132 |
| abstract_inverted_index.repair | 120 |
| abstract_inverted_index.report | 88 |
| abstract_inverted_index.strand | 116 |
| abstract_inverted_index.target | 46 |
| abstract_inverted_index.alleles | 147 |
| abstract_inverted_index.binding | 70 |
| abstract_inverted_index.defined | 13 |
| abstract_inverted_index.elicits | 115 |
| abstract_inverted_index.family, | 80 |
| abstract_inverted_index.genomes | 150 |
| abstract_inverted_index.methods | 161 |
| abstract_inverted_index.similar | 31 |
| abstract_inverted_index.system, | 110, 167 |
| abstract_inverted_index.ErCas12a | 93 |
| abstract_inverted_index.Further, | 125 |
| abstract_inverted_index.However, | 56 |
| abstract_inverted_index.activity | 94, 102 |
| abstract_inverted_index.adjacent | 63 |
| abstract_inverted_index.applying | 175 |
| abstract_inverted_index.effector | 3, 58 |
| abstract_inverted_index.mediated | 118 |
| abstract_inverted_index.nuclease | 66 |
| abstract_inverted_index.proteins | 4, 59 |
| abstract_inverted_index.provides | 160 |
| abstract_inverted_index.reported | 101, 129 |
| abstract_inverted_index.reporter | 109, 146 |
| abstract_inverted_index.sequence | 41 |
| abstract_inverted_index.specific | 22 |
| abstract_inverted_index.utilizes | 134 |
| abstract_inverted_index.variable | 18 |
| abstract_inverted_index.ErCas12a, | 73 |
| abstract_inverted_index.GeneWeld, | 137 |
| abstract_inverted_index.Recently, | 72 |
| abstract_inverted_index.Together, | 157 |
| abstract_inverted_index.annealing | 117 |
| abstract_inverted_index.deploying | 163 |
| abstract_inverted_index.effector. | 25, 55 |
| abstract_inverted_index.generally | 30 |
| abstract_inverted_index.homology, | 136 |
| abstract_inverted_index.integrate | 145 |
| abstract_inverted_index.kinetics. | 71 |
| abstract_inverted_index.secondary | 50 |
| abstract_inverted_index.structure | 51 |
| abstract_inverted_index.targeting | 7, 131 |
| abstract_inverted_index.zebrafish | 96, 153 |
| abstract_inverted_index.CRISPR-Cas | 2, 166 |
| abstract_inverted_index.additional | 165 |
| abstract_inverted_index.consisting | 36 |
| abstract_inverted_index.identified | 82 |
| abstract_inverted_index.increasing | 169 |
| abstract_inverted_index.nucleotide | 40 |
| abstract_inverted_index.previously | 100, 128 |
| abstract_inverted_index.recognized | 52 |
| abstract_inverted_index.activities, | 67 |
| abstract_inverted_index.demonstrate | 139 |
| abstract_inverted_index.efficiently | 122 |
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| abstract_inverted_index.fluorescent | 108 |
| abstract_inverted_index.protospacer | 62 |
| abstract_inverted_index.researchers | 172 |
| abstract_inverted_index.CRISPR-Cas9. | 124 |
| abstract_inverted_index.rectale</i>. | 85 |
| abstract_inverted_index.complementary | 42 |
| abstract_inverted_index.double-strand | 10 |
| abstract_inverted_index.requirements, | 65 |
| abstract_inverted_index.technologies. | 178 |
| abstract_inverted_index.<i>Eubacterium | 84 |
| abstract_inverted_index.CRISPR-ErCas12a | 114, 143 |
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| cited_by_percentile_year.max | 99 |
| cited_by_percentile_year.min | 98 |
| corresponding_author_ids | https://openalex.org/A5023581104, https://openalex.org/A5044571120 |
| countries_distinct_count | 1 |
| institutions_distinct_count | 15 |
| corresponding_institution_ids | https://openalex.org/I1330342723, https://openalex.org/I173911158 |
| citation_normalized_percentile.value | 0.92063145 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | True |