RNA interference may result in unexpected phenotypes in Caenorhabditis elegans

Author(s):Show less -

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]}

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

• 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: 12/00195-3 - Post-transcriptional regulatory networks mediated by RNA binding proteins in human pluripotent stem cells
• Grantee: Katlin Brauer Massirer
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 10/52557-0 - Identification of mechanisms responsible for beneficial effects of calorie restriction
• Grantee: Marcelo Alves da Silva Mori
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 17/22057-5 - Physiological effects and transgenerational inheritance induced by cholesterol-rich diet in Caenorhabditis elegans
• Grantee: Willian Goulart Salgueiro
• Support Opportunities: 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 Opportunities: 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 Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 16/02207-0 - Identification of soma-germline interactions that may influence lifespan in Caenorhabditis elegans
• Grantee: Evandro Araújo de Souza
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• 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 Opportunities: Scholarships in Brazil - Master
• 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 Opportunities: Regular Research Grants
• FAPESP's process: 17/01339-2 - Uncovering mechanisms of longevity mediated by mobile RNAs
• Grantee: Henrique Camara
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 14/10814-8 - Determination of factors that controls Dicer expression in vivo in C. elegans
• Grantee: Evandro Araújo de Souza
• Support Opportunities: 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 Opportunities: Research Projects - Thematic 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 Opportunities: Scholarships in Brazil - Master
• 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 Opportunities: Scholarships in Brazil - Master