CRISPR-Induced Expression of N-Terminally Truncated Dicer in Mouse Cells

0555394 - ÚMG 2022 RIV CH eng J - Článek v odborném periodiku
Malík, Radek - Svoboda, Petr
CRISPR-Induced Expression of N-Terminally Truncated Dicer in Mouse Cells.
Genes. Roč. 12, č. 4 (2021), č. článku 540. E-ISSN 2073-4425
Grant CEP: GA ČR GX20-03950X
Institucionální podpora: RVO:68378050
Klíčová slova: RNAi * Dicer * crispr * dCas9 * vp64 * ms2 * sgRNA
Obor OECD: Genetics and heredity (medical genetics to be 3)
Impakt faktor: 4.141, rok: 2021
Způsob publikování: Open access
https://www.mdpi.com/2073-4425/12/4/540

RNA interference (RNAi) designates sequence-specific mRNA degradation mediated by small RNAs generated from long double-stranded RNA (dsRNA) by RNase III Dicer. RNAi appears inactive in mammalian cells except for mouse oocytes, where high RNAi activity exists because of an N-terminally truncated Dicer isoform, denoted Dicer(O). Dicer(O) processes dsRNA into small RNAs more efficiently than the full-length Dicer expressed in somatic cells. Dicer(O) is expressed from an oocyte-specific promoter of retrotransposon origin, which is silenced in other cell types. In this work, we evaluated CRISPR-based strategies for epigenetic targeting of the endogenous Dicer gene to restore Dicer(O) expression and, consequently, RNAi. We show that reactivation of Dicer(O) expression can be achieved in mouse embryonic stem cells, but it is not sufficient to establish a robust canonical RNAi response.
Trvalý link: http://hdl.handle.net/11104/0329915