Evaluation of the Collateral Activity of CRISPR/Cas13a-Ribonuclease on the Agilent 2100 Bioanalyzer

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Published: Jun 30, 2022
DOI: https://doi.org/10.18097/BMCRM00169
Keywords:
CRISPR/Cas13a-ribonuclease; collateral activity; detection; Agilent 2100 bioanalyzer

Main Article Content

L.K. Kurbatov
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
S.P. Radko
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
S.A. Khmeleva
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
O.S. Timoshenko
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
A.V. Lisitsa
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia

Abstract

The paper describes an original technique for detecting the collateral activity of CRISPR/Cas 13a ribonuclease based on the assessment of ribosomal RNA degradation. The Agilent 2100 bioanalyzer is used as an analyzing device. This approach is an alternative to existing detection methods and has a number of advantages over them in the case when a quantitative assessment of activity is not required. On the example of the test sample, the optimal concentrations and ratios of the components of the reaction mixture, which are necessary to obtain the most indicative result, were determined. The proposed technique can be used for qualitative assessment of the activity of recombinant ribonuclease Cas13a preparations obtained under different conditions of heterologous protein expression and purification, as well as for testing guide RNAs.

Article Details

How to Cite
Kurbatov, L., Radko, S. ., Khmeleva, S., Timoshenko, O., & Lisitsa, A. (2022). Evaluation of the Collateral Activity of CRISPR/Cas13a-Ribonuclease on the Agilent 2100 Bioanalyzer. Biomedical Chemistry: Research and Methods, 5(2), e00169. https://doi.org/10.18097/BMCRM00169
Issue
Vol. 5 No. 2 (2022)
Section
PROTOCOLS OF EXPERIMENTS, USEFUL MODELS, PROGRAMS AND SERVICES

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