{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,10,19]],"date-time":"2024-10-19T04:04:48Z","timestamp":1729310688947,"version":"3.27.0"},"reference-count":23,"publisher":"The Russian Academy of Sciences","issue":"2","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molekul\u00e2rna\u00e2 biologi\u00e2"],"abstract":"<jats:p>In the paper, we propose an RNA interference-based method of inducible knockdown of genes essential for cell viability. The method arranges a genetic cassette in which an inducible metallothionein promoter controls the expression of siRNA precursor. The cassette is inserted into the genomic pre-integrated transgene by CRIPSR-Cas9. The expression of siRNA precursor and following silencing of the gene of interest is activated by the supplementation of the medium with copper ions. This technique with the production of endogenous siRNAs allows the gene knockdown in cell cultures that are refractory to conventional transfection strategies of exogenous siRNA. The efficiency of the developed method was demonstrated in the cell culture of Drosophila ovarian somatic cells 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 the differentiation of the ovarian somatic cells.<\/jats:p>","DOI":"10.31857\/s0026898424020137","type":"journal-article","created":{"date-parts":[[2024,10,9]],"date-time":"2024-10-09T14:45:21Z","timestamp":1728485121000},"page":"305-313","source":"Crossref","is-referenced-by-count":0,"title":["Method of inducible knockdown of essential genes in osc cell culture of &lt;i&gt;Drosophila melanogaster&lt;\/i&gt;"],"prefix":"10.31857","volume":"58","author":[{"given":"S. V.","family":"Marfina","sequence":"additional","affiliation":[{"name":"National Research Center \u201cKurchatov Institute\u201d"},{"name":"Mendeleev University of Chemical Technology"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E. A.","family":"Mikhaleva","sequence":"additional","affiliation":[{"name":"National Research Center \u201cKurchatov Institute\u201d"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"N. V.","family":"Akulenko","sequence":"additional","affiliation":[{"name":"National Research Center \u201cKurchatov Institute\u201d"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S. S.","family":"Ryazansky","sequence":"additional","affiliation":[{"name":"National Research Center \u201cKurchatov Institute\u201d"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"17106","published-online":{"date-parts":[[2024,10,9]]},"reference":[{"key":"ref1","doi-asserted-by":"crossref","unstructured":"Cong L., Ran F.A., Cox D., Lin S., Barretto R., Habib N., Hsu P.D., Wu X., Jiang W., Marraffini L.A., Zhang F. (2013) Multiplex genome engineering using CRISPR\/Cas Systems. Science. 339, 819\u2013823.","DOI":"10.1126\/science.1231143"},{"key":"ref2","doi-asserted-by":"crossref","unstructured":"Mali P., Yang L., Esvelt K.M., Aach J., Guell M., DiCarlo J.E., Norville J.E., Church G.M. (2013) RNA-guided human genome engineering via Cas9. Science. 339, 823\u2013826.","DOI":"10.1126\/science.1232033"},{"key":"ref3","doi-asserted-by":"crossref","unstructured":"Mohr S., Smith J., Shamu C., Neum\u00fcller R., Perrimon N. (2014) RNAi screening comes of age: improved techniques and complementary approaches. Nat. Rev. Mol. Cell. Biol. 15, 591\u2013600.","DOI":"10.1038\/nrm3860"},{"key":"ref4","doi-asserted-by":"crossref","unstructured":"Ni J.-Q., Zhou R., Czech B., Liu L.P., Holderbaum L., Yang-Zhou D., Shim H.S., Tao R., Handler D., Karpowicz P., Binari R., Booker M., Brennecke J., Perkins L.A., Hannon G.J., Perrimon N. (2011) A genome-scale shRNA resource for transgenic RNAi in Drosophila. Nat. Methods. 8, 405\u2013407.","DOI":"10.1038\/nmeth.1592"},{"key":"ref5","doi-asserted-by":"crossref","unstructured":"Gilbert L.A., Larson M.H., Morsut L., Liu Z., Brar G.A., Torres S.E., Stern-Ginossar N., Brandman O., Whitehead E.H., Doudna J.A., Lim W.A., Weissman J.S., Qi L.S. (2013) CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes. Cell. 154, 442\u2013451.","DOI":"10.1016\/j.cell.2013.06.044"},{"key":"ref6","doi-asserted-by":"crossref","unstructured":"Xu X., Qi L.S. (2019) A CRISPR\u2013dCas toolbox for genetic engineering and synthetic biology. J. Mol. Biol. 431, 34\u201347.","DOI":"10.1016\/j.jmb.2018.06.037"},{"key":"ref7","doi-asserted-by":"crossref","unstructured":"Kordy\u015b M., Sen R., Warkocki Z. (2022) Applications of the versatile CRISPR-Cas13 RNA targeting system. WIREs RNA. 13, e1694.","DOI":"10.1002\/wrna.1694"},{"key":"ref8","doi-asserted-by":"crossref","unstructured":"Scharf I., Bierbaumer L., Huber H., Wittmann P., Haider C., Pirker C., Berger W., Mikulits W. (2018) Dynamics of CRISPR\/Cas9-mediated genomic editing of the AXL locus in hepatocellular carcinoma cells. Oncol. Lett. 15, 2441\u20132450.","DOI":"10.3892\/ol.2017.7605"},{"key":"ref9","doi-asserted-by":"crossref","unstructured":"Boettcher M., McManus M.T. (2015) Choosing the right tool for the job: RNAi, TALEN, or CRISPR. Mol. Cell. 58, 575\u2013585.","DOI":"10.1016\/j.molcel.2015.04.028"},{"key":"ref10","doi-asserted-by":"crossref","unstructured":"Housden B.E., Muhar M., Gemberling M., Gersbach C.A., Stainier D.Y.R., Seydoux G., Mohr S.E., Zuber J., Perrimon N. (2017) Loss-of-function genetic tools for animal models: cross-species and cross-platform differences. Nat. Rev. Genet. 18, 24\u201340.","DOI":"10.1038\/nrg.2016.118"},{"key":"ref11","doi-asserted-by":"crossref","unstructured":"Chong Z.X., Yeap S.K., Ho W.Y. (2021) Transfection types, methods and strategies: a technical review. Peer J. 9, e11165.","DOI":"10.7717\/peerj.11165"},{"key":"ref12","doi-asserted-by":"crossref","unstructured":"Harper J.W., Schulman B.A. (2021) Cullin-RING ubiquitin ligase regulatory circuits: a quarter century beyond the F-box hypothesis. Annu. Rev. Biochem. 90, 403\u2012429.","DOI":"10.1146\/annurev-biochem-090120-013613"},{"key":"ref13","doi-asserted-by":"crossref","unstructured":"Sun J., Deng W.-M. (2005) Notch-dependent downregulation of the homeodomain gene cut is required for the mitotic cycle\/endocycle switch and cell differentiation in Drosophila follicle cells. Development. 132, 4299\u20134308.","DOI":"10.1242\/dev.02015"},{"key":"ref14","doi-asserted-by":"crossref","unstructured":"Knapp E.M., Li W., Sun J. (2019) Downregulation of homeodomain protein Cut is essential for Drosophila follicle maturation and ovulation. Development. 146, dev179002.","DOI":"10.1242\/dev.179002"},{"key":"ref15","doi-asserted-by":"crossref","unstructured":"Bassett A.R., Tibbit C., Ponting C.P., Liu J.-L. (2014) Mutagenesis and homologous recombination in Drosophila cell lines using CRISPR\/Cas9. Biol. Open. 3, 42\u201349.","DOI":"10.1242\/bio.20137120"},{"key":"ref16","doi-asserted-by":"crossref","unstructured":"Niki Y., Yamaguchi T., Mahowald A.P. (2006) Establishment of stable cell lines of Drosophila germ-line stem cells. Proc. Natl. Acad. Sci. USA. 103, 16325\u201316330.","DOI":"10.1073\/pnas.0607435103"},{"key":"ref17","doi-asserted-by":"crossref","unstructured":"Gratz S.J., Ukken F.P., Rubinstein C.D., Thiede G., Donohue L.K., Cummings A.M., O\ua78cConnor-Giles K.M. (2014) Highly specific and efficient CRISPR\/Cas9-catalyzed homology-directed repair in Drosophila. Genetics. 196, 961\u2013971.","DOI":"10.1534\/genetics.113.160713"},{"key":"ref18","doi-asserted-by":"crossref","unstructured":"Kunzelmann S., B\u00f6ttcher R., Schmidts I., F\u00f6rstemann K. (2016) A comprehensive toolbox for genome editing in cultured Drosophila melanogaster cells. G3: Genes, Genomes, Genetics. 6, 1777\u20131785.","DOI":"10.1534\/g3.116.028241"},{"key":"ref19","doi-asserted-by":"crossref","unstructured":"B\u00f6ttcher R., Hollmann M., Merk K., Nitschko V., Obermaier C., Philippou-Massier J., Wieland I., Gaul U., F\u00f6rstemann K. (2014) Efficient chromosomal gene modification with CRISPR\/cas9 and PCR-based homologous recombination donors in cultured Drosophila cells. Nucl. Acids Res. 42, e89\u2013e89.","DOI":"10.1093\/nar\/gku289"},{"key":"ref20","doi-asserted-by":"crossref","unstructured":"Perkins L.A., Holderbaum L., Tao R., Hu Y., Sopko R., McCall K., Yang-Zhou D., Flockhart I., Binari R., Shim H.S., Miller A., Housden A., Foos M., Randkelv S., Kelley C., Namgyal P., Villalta C., Liu L.P., Jiang X., Huan-Huan Q., Wang X., Fujiyama A., Toyoda A., Ayers K., Blum A., Czech B., Neumuller R., Yan D., Cavallaro A., Hibbard K., Hall D., Cooley L., Hannon G.J., Lehmann R., Parks A., Mohr S.E., Ueda R., Kondo S., Ni J.Q., Perrimon N. (2015) The transgenic RNAi project at Harvard medical school: resources and validation. Genetics. 201, 843\u2013852.","DOI":"10.1534\/genetics.115.180208"},{"key":"ref21","doi-asserted-by":"crossref","unstructured":"Sytnikova Y.A., Rahman R., Chirn G.-W., Clark J.P., Lau N.C. (2014) Transposable element dynamics and PIWI regulation impacts lncRNA and gene expression diversity in Drosophila ovarian cell cultures. Genome Res. 24, 1977\u20131990.","DOI":"10.1101\/gr.178129.114"},{"key":"ref22","doi-asserted-by":"crossref","unstructured":"Stoyko D., O T., Hernandez A., Konstantinidou P., Meng Q., Haase A.D. (2022) CRISPR-Cas9 genome editing and rapid selection of cell pools. Curr. Protoc. 2, e624.","DOI":"10.1002\/cpz1.624"},{"key":"ref23","doi-asserted-by":"crossref","unstructured":"Xia B., Amador G., Viswanatha R., Zirin J., Mohr S.E., Perrimon N. (2020) CRISPR-based engineering of gene knockout cells by homology-directed insertion in polyploid Drosophila S2R+ cells. Nat. 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