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RESEARCH PRODUCT

A Comparison of Techniques to Evaluate the Effectiveness of Genome Editing

Yegor S. VassetzkyYegor S. VassetzkyM. LipinskiTatiana TsfasmanNikolajs SjaksteDiego GerminiVlada V. Zakharova

subject

0301 basic medicineDNA End-Joining Repair[SDV.BIO]Life Sciences [q-bio]/BiotechnologyBioengineeringComputational biologyBiologyDNA sequencing03 medical and health sciencesGenome editingScreening methodAnimalsHumansDNA Breaks Double-StrandedHomologous RecombinationComputingMilieux_MISCELLANEOUSGeneticsGene EditingHigh-Throughput Nucleotide SequencingPlantsEndonucleasesZinc finger nuclease030104 developmental biologyCRISPR-Cas SystemsGenetic EngineeringBiotechnologyRNA Guide Kinetoplastida

description

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.

10.1016/j.tibtech.2017.10.008https://hal.archives-ouvertes.fr/hal-02323152