Universität zu Köln

Stankovic, Dimitrije, Csordas, Gabor ORCID: 0000-0001-6871-6839 and Uhlirova, Mirka (2022). Drosophila pVALIUM10 TRiP RNAi lines cause undesired silencing of Gateway-based transgenes. Life Sci. Alliance, 6 (2). COLD SPRING HARBOR: LIFE SCIENCE ALLIANCE LLC. ISSN 2575-1077

Full text not available from this repository.

Abstract

Post-transcriptional gene silencing using double-stranded RNA has revolutionized the field of functional genetics, allowing fast and easy disruption of gene function in various organisms. In Drosophila, many transgenic RNAi lines have been generated in large-scale efforts, including the Drosophila Transgenic RNAi Project (TRiP), to facilitate in vivo knockdown of virtually any Drosophila gene with spatial and temporal resolution. The available transgenic RNAi lines represent a fundamental resource for the fly community, providing an unprecedented opportunity to address a vast range of biological questions relevant to basic and biomedical research fields. However, caution should be applied regarding the efficiency and specificity of the RNAi approach. Here, we demonstrate that pVALIUM10-based RNAi lines, representing similar to 13% of the total TRiP collection (1,808 of 13,410 pVALIUM TRiP-based RNAi lines), cause unintended off-target silencing of transgenes expressed from Gateway destination vectors. The silencing is mediated by targeting attB1 and attB2 sequences generated via site-specific recombination and included in the transcribed mRNA. Deleting these attB sites from the Gateway expression vector prevents silencing and restores expected transgene expression.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Stankovic, Dimitrije UNSPECIFIED UNSPECIFIED UNSPECIFIED Csordas, Gabor UNSPECIFIED orcid.org/0000-0001-6871-6839 UNSPECIFIED Uhlirova, Mirka UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-662412
DOI:	10.26508/lsa.202201801
Journal or Publication Title:	Life Sci. Alliance
Volume:	6
Number:	2
Date:	2022
Publisher:	LIFE SCIENCE ALLIANCE LLC
Place of Publication:	COLD SPRING HARBOR
ISSN:	2575-1077
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DOUBLE-STRANDED-RNA; GENE-FUNCTION; R-LOOPS; SCREEN; INTERFERENCE; REGULATORS; VIVO; COLLECTION; LIBRARY; PROJECT Multiple languages Biology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/66241