Optimized knock-in of point mutations in zebrafish using CRISPR/Cas9.
Optimized knock-in of point mutations in zebrafish using CRISPR/Cas9.
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DOI：
10.1093/nar/gky512
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发表时间：
2018-09-28
影响因子：
14.9
通讯作者：
Berman JN
中科院分区：
文献类型：
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作者：
Prykhozhij SV;Fuller C;Steele SL;Veinotte CJ;Razaghi B;Robitaille JM;McMaster CR;Shlien A;Malkin D;Berman JN
We have optimized point mutation knock-ins into zebrafish genomic sites using clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 reagents and single-stranded oligodeoxynucleotides. The efficiency of knock-ins was assessed by a novel application of allele-specific polymerase chain reaction and confirmed by high-throughput sequencing. Anti-sense asymmetric oligo design was found to be the most successful optimization strategy. However, cut site proximity to the mutation and phosphorothioate oligo modifications also greatly improved knock-in efficiency. A previously unrecognized risk of off-target trans knock-ins was identified that we obviated through the development of a workflow for correct knock-in detection. Together these strategies greatly facilitate the study of human genetic diseases in zebrafish, with additional applicability to enhance CRISPR-based approaches in other animal model systems.
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影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
4.1
作者：
Liang, Xiquan;Potter, Jason;Chesnut, Jonathan D.
通讯作者：
Chesnut, Jonathan D.
DOI：
10.1073/pnas.1308335110
发表时间：
2013-08-20
影响因子：
11.1
作者：
Jao, Li-En;Wente, Susan R.;Chen, Wenbiao
通讯作者：
Chen, Wenbiao
DOI：
10.1093/dnares/dss023
发表时间：
2012-10
期刊：
DNA research : an international journal for rapid publication of reports on genes and genomes
影响因子：
--
作者：
Chen J;Zhang X;Wang T;Li Z;Guan G;Hong Y
通讯作者：
Hong Y
影响因子：
64.8
作者：
通讯作者：
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