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Vol 58(2024) N 2 p. 311-318; DOI 10.1134/S0026893324020110 Full Text

S.V. Martina1,2, E.A. Mikhaleva1, N.V. Akulenko1*, S.S. Ryazansky1**

Method of Inducible Knockdown of Essential Genes in OSC Cell Culture of Drosophila melanogaster

1Kurchatov Institute National Research Center, Moscow, 123182 Russia
2Mendeleev University of Chemical Technology, Moscow, 125047 Russia


*n.akulenko11@gmail.com
**s.ryazansky@gmail.com
Received - 2023-11-10; Revised - 2023-11-10; Accepted - 2023-11-21

An RNA interference-based method was proposed to achieve an inducible knockdown of genes essential for cell viability. In the method, a genetic cassette in which a copper ion-dependent inducible metallothionein promoter controls expression of a siRNA precursor is inserted into a genomic pre-integrated trans-gene by CRIPSR/Cas9 technology. The endogenous siRNA source allows the gene knockdown in cell cultures that are refractory to conventional transfection with exogenous siRNA. The efficiency of the method was demonstrated in Drosophila ovarian somatic cell culture (OSC) for two genes that are essential for oogenesis: Cul3, encoding a component of the multiprotein ubiquitin-ligase complex with versatile functions in proteostasis, and cut, encoding a transcription factor regulating differentiation of ovarian follicular cells.

knockdown, siRNA, CRISPR/Cas9, Drosophila, OSC



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