Construction of a germline-specific RNAi tool in C. elegans

Scientific Reports
Lina ZouDi Chen

Abstract

Analysis of complex biological functions usually requires tissue-specific genetic manipulations in multicellular organisms. The C. elegans germline plays regulatory roles not only in reproduction, but also in metabolism, stress response and ageing. Previous studies have used mutants of rrf-1, which encodes an RNA-directed RNA polymerase, as a germline-specific RNAi tool. However, the rrf-1 mutants showed RNAi activities in somatic tissues. Here we constructed a germline-specific RNAi strain by combining an indel mutation of rde-1, which encodes an Argonaute protein that functions cell autonomously to ensure RNAi efficiency, and a single copy rde-1 transgene driven by the sun-1 germline-specific promoter. The germline RNAi efficiency and specificity are confirmed by RNAi phenocopy of known mutations, knockdown of GFP reporter expression, as well as quantitative RT-PCR measurement of tissue-specific mRNAs upon RNAi knockdown. The germline-specific RNAi strain shows no obvious deficiencies in reproduction, lipid accumulation, thermo-tolerance and life span compared to wild-type animals. By screening an RNAi sub-library of phosphatase genes, we identified novel regulators of thermo-tolerance. Together, we have created a useful tool...Continue Reading

References

Aug 1, 1995·Proceedings of the National Academy of Sciences of the United States of America·G J LithgowT E Johnson
Nov 6, 1998·Nature·L Timmons, A Fire
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
May 17, 2002·Current Biology : CB·A Craig MackinnonBenjamin D Williams
Aug 19, 2006·Science·Julie M PinkstonCynthia Kenyon
Aug 8, 2007·Gene·Hiroshi QadotaKozo Kaibuchi
Dec 1, 2007·WormBook : the Online Review of C. Elegans Biology·Judith Kimble, Sarah L Crittenden
Feb 1, 2008·Development·Caroline A SpikeSusan Strome
Oct 28, 2008·Nature Genetics·Christian Frøkjaer-JensenErik M Jorgensen
Jan 27, 2009·Proceedings of the National Academy of Sciences of the United States of America·Antony M JoseCraig P Hunter
Nov 4, 2009·Cell Metabolism·Eyleen J O'RourkeGary Ruvkun
Feb 24, 2010·Proceedings of the National Academy of Sciences of the United States of America·Shaobing O ZhangHo Yi Mak
Jun 1, 2010·Nature Methods·Andrea CalixtoMartin Chalfie
Nov 3, 2016·Proceedings of the National Academy of Sciences of the United States of America·Julia MarréAntony M Jose

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Citations

Sep 19, 2020·G3 : Genes - Genomes - Genetics·Jason S WattsJennifer L Watts
Oct 2, 2020·Ecotoxicology and Environmental Safety·Yunhan YangDayong Wang
Jun 17, 2020·Developmental Cell·Nicole M TemplemanColeen T Murphy
Apr 24, 2021·Ecotoxicology and Environmental Safety·Shuting WangDayong Wang

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Methods Mentioned

BETA
transgenic
gene knockdown
PCR

Software Mentioned

Adobe Photoshop
GraphPad Prism
ImageJ

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