Out Knocker: a web tool for rapid and simple genotyping of designer nuclease edited cell lines

被引：106

作者：

Schmid-Burgk, Jonathan L. ^{[1
]}

Schmidt, Tobias ^{[1
]}

Gaidt, Moritz M. ^{[1
]}

Pelka, Karin ^{[2
]}

Latz, Eicke ^{[2
]}

Ebert, Thomas S. ^{[1
]}

Hornung, Veit ^{[1
]}

机构：

[1] Univ Bonn, Univ Hosp, Inst Mol Med, D-53127 Bonn, Germany

[2] Univ Bonn, Univ Hosp, Inst Innate Immun, D-53127 Bonn, Germany

来源：

GENOME RESEARCH | 2014年 / 24卷 / 10期

基金：

欧洲研究理事会;

关键词：

ZINC-FINGER NUCLEASES; GENOME; IMMUNITY;

D O I：

10.1101/gr.176701.114

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The application of designer nucleases allows the induction of DNA double-strand breaks (DSBs) at user-defined genomic loci. Due to imperfect DNA repair mechanisms, DSBs can lead to alterations in the genomic architecture, such as the disruption of the reading frame of a critical exon. This can be exploited to generate somatic knockout cell lines. While high genome editing activities can be achieved in various cellular systems, obtaining cell clones that contain all-allelic frameshift mutations at the target locus of interest remains a laborious task. To this end, we have developed an easy-to-follow deep sequencing workflow and the evaluation tool OutKnocker (www.OutKnocker.org), which allows convenient, reliable, and cost-effective identification of knockout cell lines.

引用

页码：1719 / 1723

页数：5

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