CRISPR Interference of Adenylate Cyclases from Mycobacterium tuberculosis

N I Nadolinskaia; M V Zamakhaev; M S Shumkov; D K Armianinova; D S Karpov; A V Goncharenko

doi:10.1134/S0003683821040128

CRISPR Interference of Adenylate Cyclases from Mycobacterium tuberculosis

Appl Biochem Microbiol. 2021;57(4):421-425. doi: 10.1134/S0003683821040128. Epub 2021 Jul 27.

Authors

N I Nadolinskaia¹, M V Zamakhaev¹, M S Shumkov¹, D K Armianinova¹, D S Karpov², A V Goncharenko¹

Affiliations

¹ Bach Institute of Biochemistry, Fundamentals of Biotechnology Federal Research Center, Russian Academy of Sciences, 119071 Moscow, Russia.
² Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia.

Abstract

This work describes a modification of the pRH2521 vector of the pRH2502/pRH2521 system for CRISPR-dCas9-mediated RNA interference. The modification enabled an increase in the cloning efficiency of guide RNA spacers. The ability of the modified pRH2502/pRH2521 system to suppress the transcription of certain genes was evaluated with the use of genes of Mycobacterium tuberculosis adenylate cyclases. The results revealed the limitations of the pRH2502/pRH2521 system for CRISPR interference associated with the probability of the detection of a protospacer adjacent motif (PAM) in the gene promoter region.

Keywords: CRISPR; CRISPRi; adenylate cyclase; mycobacteria; tuberculosis.

© Pleiades Publishing, Inc. 2021, ISSN 0003-6838, Applied Biochemistry and Microbiology, 2021, Vol. 57, No. 4, pp. 421–425. © Pleiades Publishing, Inc., 2021.Russian Text © The Author(s), 2021, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2021, Vol. 57, No. 4, pp. 326–331.