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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

RNA interference may result in unexpected phenotypes in Caenorhabditis elegans

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Author(s):
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De-Souza, Evandro A. [1, 2] ; Camara, Henrique [3, 4, 1] ; Salgueiro, Willian G. [3, 4] ; Moro, Raissa P. [3, 4] ; Knittel, Thiago L. [3, 4] ; Tonon, Guilherme [3, 4] ; Pinto, Silas [3, 4, 1] ; Pinca, Ana Paula F. [1] ; Antebi, Adam [5, 6] ; Pasquinelli, Amy E. [7] ; Massirer, Katlin B. [4, 8, 9] ; Mori, Marcelo A. [3, 4, 1]
Total Authors: 12
Affiliation:
[1] Univ Fed Sao Paulo, Program Mol Biol, BR-04044020 Sao Paulo - Brazil
[2] Univ Fed Rio de Janeiro, Inst Bioquim Med Leopoldo de Meis, Program Mol Biol & Biotechnol, BR-21941902 Rio De Janeiro - Brazil
[3] Univ Estadual Campinas, Dept Biochem & Tissue Biol, BR-13083862 Campinas, SP - Brazil
[4] Univ Estadual Campinas, Program Genet & Mol Biol, BR-13083970 Campinas, SP - Brazil
[5] Max Planck Inst Biol Ageing, D-50931 Cologne - Germany
[6] Univ Cologne, Cologne Excellence Cluster Cellular Stress Respon, D-50931 Cologne - Germany
[7] Univ Calif San Diego, Div Biol, La Jolla, CA 92093 - USA
[8] Univ Estadual Campinas, CBMEG UNICAMP, Ctr Mol Biol & Genet Engn, BR-13083875 Campinas, SP - Brazil
[9] Univ Estadual Campinas, Struct Genom Consortium UNICAMP, BR-13083875 Campinas, SP - Brazil
Total Affiliations: 9
Document type: Journal article
Source: Nucleic Acids Research; v. 47, n. 8, p. 3957-3969, MAY 7 2019.
Web of Science Citations: 3
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. (AU)

FAPESP's process: 17/22057-5 - Physiological effects and transgenerational inheritance induced by cholesterol-rich diet in Caenorhabditis elegans
Grantee:Willian Goulart Salgueiro
Support type: Scholarships in Brazil - Master
FAPESP's process: 14/10814-8 - Determination of factors that controls Dicer expression in vivo in C. elegans
Grantee:Evandro Araújo de Souza
Support type: Scholarships in Brazil - Master
FAPESP's process: 17/08829-5 - Investigation of the interaction between Dicer and the NAD+ signalling pathway in C. elegans metabolism and aging
Grantee:Guilherme Tonon da Silva
Support type: Scholarships in Brazil - Master
FAPESP's process: 17/01184-9 - CAMeLEOm: cross-species analysis of metabolic, lifespan effects and omics of dietary restriction mimetics
Grantee:Marcelo Alves da Silva Mori
Support type: Research Projects - Thematic Grants
FAPESP's process: 15/01316-7 - Dicer, miRNAs and the control of mitochondrial function in the context of aging and caloric restriction
Grantee:Marcelo Alves da Silva Mori
Support type: Regular Research Grants
FAPESP's process: 14/25068-0 - Unraveling new mechanisms responsible for the cell non-autonomous control of proteostasis in c. elegans
Grantee:Ana Paula Forti Pinca
Support type: Scholarships in Brazil - Master
FAPESP's process: 17/01339-2 - Uncovering mechanisms of longevity mediated by mobile RNAs
Grantee:Henrique Camara
Support type: Scholarships in Brazil - Doctorate
FAPESP's process: 17/03423-0 - Characterization of the Tissue-Specific Alterations in Protein Expression with Aging and Caloric Restriction in C. Elegans
Grantee:Thiago Leite Knittel
Support type: Scholarships in Brazil - Master
FAPESP's process: 12/00195-3 - Post-transcriptional regulatory networks mediated by RNA binding proteins in human pluripotent stem cells
Grantee:Katlin Brauer Massirer
Support type: Research Grants - Young Investigators Grants
FAPESP's process: 16/02207-0 - Identification of soma-germline interactions that may influence lifespan in Caenorhabditis elegans
Grantee:Evandro Araújo de Souza
Support type: Scholarships abroad - Research Internship - Master's degree
FAPESP's process: 16/15958-3 - Soma-Germline interaction and its effect on longevity: the role of Argonautes in aging c. elegans
Grantee:Raíssa de Paula Moro
Support type: Scholarships in Brazil - Master
FAPESP's process: 17/04377-2 - Study of Dicer effects on mitochondrial function, metabolism and aging in c. elegans nematodes
Grantee:Silas Pinto da Silva
Support type: Scholarships in Brazil - Doctorate