Expanding the CRISPR Toolbox with ErCas12a in Zebrafish and Human Cells

dc.contributor.author McGrail, Maura
dc.contributor.author Wierson, Wesley
dc.contributor.author Simone, Brandon
dc.contributor.author WareJoncas, Zachary
dc.contributor.author Mann, Carla
dc.contributor.author Essner, Jeffrey
dc.contributor.author Welker, Jordan
dc.contributor.author Kar, Bibekananda
dc.contributor.author Emch, Michael
dc.contributor.author Friedberg, Iddo
dc.contributor.author Gendron, William
dc.contributor.author Barry, Michael
dc.contributor.author Clark, Karl
dc.contributor.author Dobbs, Drena
dc.contributor.author McGrail, Maura
dc.contributor.author Ekker, Stephen
dc.contributor.author Essner, Jeffrey
dc.contributor.department Veterinary Microbiology and Preventive Medicine
dc.contributor.department Genetics, Development and Cell Biology
dc.date 2020-04-07T17:46:17.000
dc.date.accessioned 2020-06-30T04:02:12Z
dc.date.available 2020-06-30T04:02:12Z
dc.date.copyright Tue Jan 01 00:00:00 UTC 2019
dc.date.issued 2019-12-01
dc.description.abstract <p>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 <em>Eubacterium rectale</em>. 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.</p>
dc.description.comments <p>This article is published as Wierson, Wesley A., Brandon W. Simone, Zachary WareJoncas, Carla Mann, Jordan M. Welker, Bibekananda Kar, Michael J. Emch et al. "Expanding the CRISPR Toolbox with ErCas12a in Zebrafish and Human Cells." <em>The CRISPR journal</em> 2, no. 6 (2019): 417-433. doi: <a href="https://doi.org/10.1089/crispr.2019.0026">10.1089/crispr.2019.0026</a>.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/gdcb_las_pubs/242/
dc.identifier.articleid 1244
dc.identifier.contextkey 17230218
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath gdcb_las_pubs/242
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/37919
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/gdcb_las_pubs/242/2019_McGrail_ExpandingToolbox.pdf|||Fri Jan 14 22:53:05 UTC 2022
dc.source.uri 10.1089/crispr.2019.0026
dc.subject.disciplines Cell and Developmental Biology
dc.subject.disciplines Genetics and Genomics
dc.subject.disciplines Molecular Genetics
dc.subject.disciplines Veterinary Preventive Medicine, Epidemiology, and Public Health
dc.title Expanding the CRISPR Toolbox with ErCas12a in Zebrafish and Human Cells
dc.type article
dc.type.genre article
dspace.entity.type Publication
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