Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase
The CRISPR/Cas9 nickase mutant is less prone to off-target double-strand (ds)DNA breaks than wild-type Cas9 because to produce dsDNA cleavage it requires two guide RNAs to target the nickase to nearby opposing strands. Like wild-type Cas9 lesions, these staggered lesions are repaired by either non-h...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Oxford University Press
2019-01-01
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| Series: | G3: Genes, Genomes, Genetics |
| Subjects: | |
| Online Access: | http://g3journal.org/lookup/doi/10.1534/g3.118.200758 |
| _version_ | 1818657324009521152 |
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| author | Xi A. Ge Craig P. Hunter |
| author_facet | Xi A. Ge Craig P. Hunter |
| author_sort | Xi A. Ge |
| collection | DOAJ |
| description | The CRISPR/Cas9 nickase mutant is less prone to off-target double-strand (ds)DNA breaks than wild-type Cas9 because to produce dsDNA cleavage it requires two guide RNAs to target the nickase to nearby opposing strands. Like wild-type Cas9 lesions, these staggered lesions are repaired by either non-homologous end joining or, if a repair template is provided, by homologous recombination (HR). Here, we report very efficient (up to 100%) recovery of heterozygous insertions in Mus musculus produced by long (>300 nt), single-stranded DNA donor template-guided repair of paired-nickase lesions. |
| first_indexed | 2024-12-17T03:39:40Z |
| format | Article |
| id | doaj.art-07c1d9e919d64d67bc3ffda2c7fc6f59 |
| institution | Directory Open Access Journal |
| issn | 2160-1836 |
| language | English |
| last_indexed | 2024-12-17T03:39:40Z |
| publishDate | 2019-01-01 |
| publisher | Oxford University Press |
| record_format | Article |
| series | G3: Genes, Genomes, Genetics |
| spelling | doaj.art-07c1d9e919d64d67bc3ffda2c7fc6f592022-12-21T22:05:02ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362019-01-019128128610.1534/g3.118.20075825Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 NickaseXi A. GeCraig P. HunterThe CRISPR/Cas9 nickase mutant is less prone to off-target double-strand (ds)DNA breaks than wild-type Cas9 because to produce dsDNA cleavage it requires two guide RNAs to target the nickase to nearby opposing strands. Like wild-type Cas9 lesions, these staggered lesions are repaired by either non-homologous end joining or, if a repair template is provided, by homologous recombination (HR). Here, we report very efficient (up to 100%) recovery of heterozygous insertions in Mus musculus produced by long (>300 nt), single-stranded DNA donor template-guided repair of paired-nickase lesions.http://g3journal.org/lookup/doi/10.1534/g3.118.200758Cas9 nickaseLong single stranded DNAssODN |
| spellingShingle | Xi A. Ge Craig P. Hunter Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase G3: Genes, Genomes, Genetics Cas9 nickase Long single stranded DNA ssODN |
| title | Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase |
| title_full | Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase |
| title_fullStr | Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase |
| title_full_unstemmed | Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase |
| title_short | Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase |
| title_sort | efficient homologous recombination in mice using long single stranded dna and crispr cas9 nickase |
| topic | Cas9 nickase Long single stranded DNA ssODN |
| url | http://g3journal.org/lookup/doi/10.1534/g3.118.200758 |
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