A Comparison of Techniques to Evaluate the Effectiveness of Genome Editing

被引:27
作者
Germini, Diego [1 ,2 ]
Tsfasman, Tatiana [1 ,2 ]
Zakharova, Vlada V. [1 ,2 ]
Sjakste, Nikolajs [3 ]
Lipinski, Marc [1 ,2 ]
Vassetzky, Yegor [1 ,2 ,4 ]
机构
[1] Univ Paris Sud Paris Saclay, CNRS, Inst Gustave Roussy, UMR 8126, F-94805 Villejuif, France
[2] French Russian Joint Canc Res Lab, LIA 1066, F-94805 Villejuif, France
[3] Univ Latvia, Fac Med, Riga, Latvia
[4] Russian Acad Sci, Koltzov Inst Dev Biol, Moscow, Russia
关键词
CONFORMATIONAL POLYMORPHISM ANALYSIS; RESOLUTION MELTING ANALYSIS; ZINC-FINGER; DNA RECOGNITION; OFF-TARGET; ON-TARGET; IN-VIVO; GENERATION; MUTATION; VALIDATION;
D O I
10.1016/j.tibtech.2017.10.008
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Genome editing using engineered nucleases (meganucleases, zinc finger nucleases, transcription activator-like effector nucleases) has created many recent breakthroughs. Prescreening for efficiency and specificity is a critical step prior to using any newly designed genome editing tool for experimental purposes. The current standard screening methods of evaluation are based on DNA sequencing or use mismatch-sensitive endonucleases. They can be time-consuming and costly or lack reproducibility. Here, we review and critically compare standard techniques with those more recently developed in terms of reliability, time, cost, and ease of use.
引用
收藏
页码:147 / 159
页数:13
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