A Comparison of Techniques to Evaluate the Effectiveness of Genome Editing

Trends Biotechnol. 2018 Feb;36(2):147-159. doi: 10.1016/j.tibtech.2017.10.008. Epub 2017 Nov 17.

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.

Keywords: efficiency; evaluation; genome editing; methods.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • DNA Breaks, Double-Stranded
  • DNA End-Joining Repair
  • Endonucleases / genetics*
  • Endonucleases / metabolism
  • Gene Editing / instrumentation
  • Gene Editing / methods*
  • Genetic Engineering / instrumentation
  • Genetic Engineering / methods*
  • High-Throughput Nucleotide Sequencing / methods
  • Homologous Recombination
  • Humans
  • Plants / genetics
  • RNA, Guide, CRISPR-Cas Systems

Substances

  • Endonucleases