GeneWeld: a method for efficient targeted integration directed by short homology McGrail, Maura Welker, Jordan Almeida, Maira Essner, Jeffrey Webster, Dennis Torrie, Melanie Weiss, Trevor Vollbrecht, Macy Lan, Merrina McKeighan, Kenna MIng, Zhitao Wehmeier, Alec Mikelson, Christopher Haltom, Jeffrey Kwan, Kristen Chien, Chi-­Bin Balciunas, Darius Ekker, Stephen Clark, Karl Webber, Beau Moriarity, Branden Solin, Staci Carlson, Daniel Dobbs, Drena McGrail, Maura Essner, Jeffrey
dc.contributor.department Genetics, Development and Cell Biology 2019-02-07T21:50:23.000 2020-06-30T04:01:53Z 2020-06-30T04:01:53Z Mon Jan 01 00:00:00 UTC 2018 2018-10-03
dc.description.abstract <p>Choices for genome engineering and integration involve high efficiency with little or no target specificity or high specificity with low activity. Here, we describe a targeted integration strategy, called GeneWeld, and a vector series for gene tagging, pGTag (plasmids for Gene Tagging), which promote highly efficient and precise targeted integration in zebrafish embryos, pig fibroblasts, and human cells utilizing the CRISPR/Cas9 system. Our work demonstrates that in vivo targeting of a genomic locus of interest with CRISPR/Cas9 and a donor vector containing as little as 24 to 48 base pairs of homology directs precise and efficient knock-in when the homology arms are exposed with a double strand break in vivo. Given our results targeting multiple loci in different species, we expect the accompanying protocols, vectors, and web interface for homology arm design to help streamline gene targeting and applications in CRISPR compatible systems.</p>
dc.description.comments <p>This is a pre-print made available through bioRxiv, doi: <a href="" target="_blank">10.1101/431627</a>.</p>
dc.format.mimetype application/pdf
dc.identifier archive/
dc.identifier.articleid 1211
dc.identifier.contextkey 13677647
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath gdcb_las_pubs/207
dc.language.iso en
dc.source.bitstream archive/|||Fri Jan 14 22:27:21 UTC 2022
dc.source.uri 10.1101/431627
dc.subject.disciplines Cell and Developmental Biology
dc.subject.disciplines Genetics
dc.subject.disciplines Genomics
dc.subject.keywords CRISPR/Cas9
dc.subject.keywords knock-­in
dc.subject.keywords homology mediated-­end joining
dc.subject.keywords targeted integration
dc.subject.keywords zebrafish
dc.subject.keywords pig fibroblasts
dc.subject.keywords human K-­562 cells
dc.title GeneWeld: a method for efficient targeted integration directed by short homology
dc.type article
dc.type.genre article
dspace.entity.type Publication
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relation.isOrgUnitOfPublication 9e603b30-6443-4b8e-aff5-57de4a7e4cb2
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