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          Institute: MPI für biophysikalische Chemie     Collection: Biochemie und Zellbiologie (Prof. Klaus Weber)     Display Documents

ID: 16932.0, MPI für biophysikalische Chemie / Biochemie und Zellbiologie (Prof. Klaus Weber)
Identification of essential genes in cultured mammalian cells using small interfering RNAs
Authors:Harborth, J.; Elbashir, S. M.; Bechert, K.; Tuschl, T.; Weber, K.
Date of Publication (YYYY-MM-DD):2001-12
Title of Journal:Journal of Cell Science
Issue / Number:24
Start Page:4557
End Page:4565
Review Status:Peer-review
Audience:Not Specified
Abstract / Description:We report the first RNAi-induced phenotypes in mammalian cultured cells using RNA interference mediated by duplexes of 21-nt RNAs. The 21 gene products studied have different functions and subcellular localizations. Knockdown experiments monitored by immunofluorescence and immunoblotting show that even major cellular proteins such as actin and vimentin can be silenced efficiently. Genes were classified as essential or nonessential depending on impaired cell growth after RNA silencing. Phenotypes also involved altered cell morphology and aberrant mitotic arrest. Among the essential genes identified by RNAi for which such information was previously not available are lamin B1, lamin B2, NUP153, GAS41, ARC21, cytoplasmic dynein, the protein kinase cdk1 and both beta- and gamma-actin. Newly defined nonessential genes are emerin and zyxin. Several genes previously characterized by other methods such as knockout of murine genes are included as internal controls and gave identical results when RNAi was used. In the case of two nonessential genes (lamin A/C and zyxin) RNAi provides a recognizable phenotype. Our results complete the characterization of the mammalian nuclear lamins. While lamins A/C appear as nonessential proteins in the mouse embryo and in RNAi treated cultured cells, the two other lamins, B1 and B2, are now identified as essential proteins. Interestingly the inner nuclear membrane protein emerin, thought to be a ligand of lamin A/C, is also a nonessential protein in tissue culture cells.
Free Keywords:functional genomics; gene silencing; mammalian cells; nuclear lamins; RNA interference
Comment of the Author/Creator:Date: 2001, DEC
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Klaus Weber / 090
MPI für biophysikalische Chemie/AG Thomas Tuschl
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