A Comparison of Techniques to Evaluate the Effectiveness of Genome Editing. Issue 2 (February 2018)

• Record Type: Journal Article
• Authors: Germini, Diego; Tsfasman, Tatiana; Zakharova, Vlada V.; Sjakste, Nikolajs; Lipinski, Marс; Vassetzky, Yegor
• Is Part Of: Trends in biotechnology. Volume 36:Issue 2(2018)
• Page Start: 147
• Page End: 159
• Publication Date: 2018-02
• Subjects: efficiency -- evaluation -- genome editing -- methods; Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Genetic engineering -- Periodicals; Industrial microbiology -- Periodicals; 660.605
• DOI: 10.1016/j.tibtech.2017.10.008
• Languages: English
• ISSNs: 0167-7799

Abstract

Abstract : 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. Trends: The number of methods to assess the efficiency of genome editing is increasing very quickly. Recent techniques tend to exploit recent advances in laboratory practice (e.g., next-generation sequencing, computational analysis). Methods for genome-wide screening developed can simultaneously determine the specificity of genome editing tools and identify the presence of off-targets. While these methods are costly and time-consuming, they are indispensable when genome editing is planned to be used in vivo . The trend in in vitro research applications is to develop techniques that simplify as much as possible the first step of testing the efficiency of a newly designed genome editing tool.