REVIEW: Practical Recommendations for Improving Efficiency and Accuracy of the CRISPR/Cas9 Genome Editing System

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M. N. Karagyaur^1,2,a*, Y. P. Rubtsov², P. A. Vasiliev³, and V. A. Tkachuk^1,2

¹Lomonosov Moscow State University, Institute of Regenerative Medicine, 119192 Moscow, Russia

²Lomonosov Moscow State University, Faculty of Fundamental Medicine, 119192 Moscow, Russia

³Research Center of Medical Genetics, Russian Academy of Medical Sciences, 115478 Moscow, Russia

^* To whom correspondence should be addressed.

Received January 11, 2018

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CRISPR/Cas9 genome-editing system is a powerful, fairly accurate, and efficient tool for modifying genomic DNA. Despite obvious advantages, it is not devoid of certain drawbacks, such as propensity for introduction of additional nonspecific DNA breaks, insufficient activity against aneuploid genomes, and relative difficulty in delivering its components to cells. In this review, we focus on the difficulties that can limit the use of CRISPR/Cas9 and suggest a number of practical recommendations and information sources that will make it easier for the beginners to work with this outstanding technological achievement of the XXI century.

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KEY WORDS: genome editing, CRISPR/Cas9, NHEJ, HDR, HITI, gRNA design, HDR/HITI template design

DOI: 10.1134/S0006297918060020

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