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CRISPR/CAS9, the king of genome editing tools

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Abstract

The discovery of CRISPR/Cas9 brought a hope for having an efficient, reliable, and readily available tool for genome editing. CRISPR/Cas9 is certainly easy to use, while its efficiency and reliability remain the focus of studies. The review describes the general principles of the organization and function of Cas nucleases and a number of important issues to be considered while planning genome editing experiments with CRISPR/Cas9. The issues include evaluation of the efficiency and specificity for Cas9, sgRNA selection, Cas9 variants designed artificially, and use of homologous recombination and nonhomologous end joining in DNA editing.

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Abbreviations

NGS:

next-generation sequencing

NHEJ:

nonhomologous end joining

HDR:

homology-directed repair

DSB:

double-strand break

CRISPR:

clustered regularly-interspaced short palindromic repeats

PAM:

protospacer adjacent motif

crRNA:

CRISPR RNA

sgRNA:

single guide RNA

tracrRNA:

trans-activating crRNA

Cas:

CRISPR-associated protein

spCas9:

Streptococcus pyogenes Cas9

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Authors and Affiliations

  1. Medical Genetic Research Center, Moscow, 115478, Russia

    A. V. Bannikov & A. V. Lavrov

  2. Pirogov Russian National Research Medical University, Moscow, 117997, Russia

    A. V. Lavrov

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  1. A. V. Bannikov
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Original Russian Text © A.V. Bannikov, A.V. Lavrov, 2017, published in Molekulyarnaya Biologiya, 2017, Vol. 51, No. 4, pp. 582–594.

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Bannikov, A.V., Lavrov, A.V. CRISPR/CAS9, the king of genome editing tools. Mol Biol 51, 514–525 (2017). https://doi.org/10.1134/S0026893317040033

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