Creation of A549 and MCF7 tumor sublines with knockout of TP53 using CRISPR/Cas9
https://doi.org/10.25557/2073-7998.2023.11.27-34
Abstract
The tumor suppressor p53 is the central point of cellular defense against oncogenic transformation. Mutations of TP53 gene are present in approximately half of human tumors and promote not only tumor progression, but also resistance to anticancer drugs. Creation of isogenic models, differing in their TP53 status, is valuable not only for studying its role in carcinogenesis, but also for screening of anticancer drugs and their combinations. Establishment of isogenic models using the CRISPR/Cas9 system is usually done through single cell cloning, which can lead to clonal effects, because of heterogeneity of the cell cultures. In this article we present the process of creation of new knockout cell sublines of MCF7 and A549 using CRISPR/Cas9 and selection using nutlin-3, which allows selecting cell sublines without clonal selection. Phenotypically the knockout of TP53 was confirmed by total absence of p53, absence of induction of p53-dependent gene CDKN1A, and shift in sensitivity to DNA-damaging drugs.
Supporting agencies: The study was supported by the Russian Science Foundation grant No. 22-24-00990, https://rscf.ru/project/22-24-00990/.
About the Authors
A. I. Khamidullina
Institute of Gene Biology Russian Academy of Sciences
Russian Federation
Russian Federation
34/5 Vavilova st., Moscow, 119334
E. R. Gandalipov
Solution Chemistry of Advanced Materials and Technologies, ITMO University
Russian Federation
Russian Federation
49, bldg. A, Kronverksky Pr., St. Petersburg, 197101
Y. E. Abramenko
Institute of Gene Biology Russian Academy of Sciences
Russian Federation
Russian Federation
34/5 Vavilova st., Moscow, 119334
K. V. Chernov
Solution Chemistry of Advanced Materials and Technologies, ITMO University
Russian Federation
Russian Federation
49, bldg. A, Kronverksky Pr., St. Petersburg, 197101
T. A. Kirukhina
Institute of Gene Biology Russian Academy of Sciences
Russian Federation
Russian Federation
34/5 Vavilova st., Moscow, 119334
A. V. Bruter
Institute of Gene Biology Russian Academy of Sciences
Russian Federation
Russian Federation
34/5 Vavilova st., Moscow, 119334
V. V. Tatarskiy
Institute of Gene Biology Russian Academy of Sciences
Russian Federation
Russian Federation
34/5 Vavilova st., Moscow, 119334
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Review
For citations:
Khamidullina A.I., Gandalipov E.R., Abramenko Y.E., Chernov K.V., Kirukhina T.A., Bruter A.V., Tatarskiy V.V. Creation of A549 and MCF7 tumor sublines with knockout of TP53 using CRISPR/Cas9. Medical Genetics. 2023;22(11):27-34. (In Russ.) https://doi.org/10.25557/2073-7998.2023.11.27-34
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