Universität zu Köln

De-Souza, Evandro A., Camara, Henrique, Salgueiro, Willian G., Moro, Raissa P., Knittel, Thiago L., Tonon, Guilherme, Pinto, Silas ORCID: 0000-0003-0894-8915, Pinca, Ana Paula F., Antebi, Adam, Pasquinelli, Amy E., Massirer, Katlin B. and Mori, Marcelo A. (2019). RNA interference may result in unexpected phenotypes in Caenorhabditis elegans. Nucleic Acids Res., 47 (8). S. 3957 - 3970. OXFORD: OXFORD UNIV PRESS. ISSN 1362-4962

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Abstract

RNA interference (RNAi) is a valuable technique to determine gene function. In Caenorhabditis elegans, RNAi can be achieved by feeding worms bacteria carrying a plasmid expressing double-stranded RNA (dsRNA) targeting a gene of interest. The most commonly used plasmid vector for this purpose is L4440. However, it has been noticed that sequences within L4440 may elicit unspecific effects. Here, we provide a comprehensive characterization of these effects and their mechanisms and describe new unexpected phenotypes uncovered by the administration of unspecific exogenous dsRNA. An example involves dsRNA produced by the multiple cloning site (MCS) of L4440, which shares complementary sequences with some widely used reporter vectors and induces partial transgene silencing via the canonical and antiviral RNAi pathway. Going beyond transgene silencing, we found that the reduced embryonic viability of mir-35-41(gk262) mutants is partially reversed by exogenous dsRNA via a mechanism that involves canonical RNAi. These results indicate cross-regulation between different small RNA pathways in C. elegans to regulate embryonic viability. Recognition of the possible unspecific effects elicited by RNAi vectors is important for rigorous interpretation of results from RNAi-based experiments.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code De-Souza, Evandro A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Camara, Henrique UNSPECIFIED UNSPECIFIED UNSPECIFIED Salgueiro, Willian G. UNSPECIFIED UNSPECIFIED UNSPECIFIED Moro, Raissa P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Knittel, Thiago L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Tonon, Guilherme UNSPECIFIED UNSPECIFIED UNSPECIFIED Pinto, Silas UNSPECIFIED orcid.org/0000-0003-0894-8915 UNSPECIFIED Pinca, Ana Paula F. UNSPECIFIED UNSPECIFIED UNSPECIFIED Antebi, Adam UNSPECIFIED UNSPECIFIED UNSPECIFIED Pasquinelli, Amy E. UNSPECIFIED UNSPECIFIED UNSPECIFIED Massirer, Katlin B. UNSPECIFIED UNSPECIFIED UNSPECIFIED Mori, Marcelo A. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-148445
DOI:	10.1093/nar/gkz154
Journal or Publication Title:	Nucleic Acids Res.
Volume:	47
Number:	8
Page Range:	S. 3957 - 3970
Date:	2019
Publisher:	OXFORD UNIV PRESS
Place of Publication:	OXFORD
ISSN:	1362-4962
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DOUBLE-STRANDED-RNA; C-ELEGANS; LIFE-SPAN; GENETIC INTERFERENCE; FUNCTIONAL-ANALYSIS; BODY-SIZE; EXPRESSION; PROTEIN; MECHANISMS; MICRORNAS Multiple languages Biochemistry & Molecular Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/14844