The CONDOR pipeline for simultaneous knockdown of multiple genes identifies RBBP-5 as a germ cell reprogramming barrier in C. elegans

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Item Type:	Preprint
Title:	The CONDOR pipeline for simultaneous knockdown of multiple genes identifies RBBP-5 as a germ cell reprogramming barrier in C. elegans
Creators Name:	Kazmierczak, M., Díaz, C.F., Ofenbauer, A. and Tursun, B.
Abstract:	Multiple gene activities control complex biological processes such as cell fate specification during development and cellular reprogramming. Investigating the manifold gene functions in biological systems requires also simultaneous depletion of two or more gene activities. RNA interference-mediated knockdown (RNAi) is commonly used in C. elegans to assess essential genes, which otherwise lead to lethality or developmental arrest upon full knockout. RNAi application is straightforward by feeding worms with RNAi plasmid-containing bacteria. However, the general approach of mixing bacterial RNAi clones to deplete two genes simultaneously often yields poor results. To address this issue, we developed a bacterial conjugation-mediated double RNAi technique ‘CONDOR’. It allows combining RNAi bacteria for robust double RNAi with high-throughput. To demonstrate the power of CONDOR for large scale double RNAi screens we conjugated RNAi against the histone chaperone gene lin-53 with more than 700 other chromatin factor genes. Thereby, we identified the Set1/MLL methyltransferase complex member RBBP-5 as a novel germ cell reprogramming barrier. Our findings demonstrate that CONDOR increases efficiency and versatility of RNAi screens to examine interconnected biological processes in C. elegans with high-throughput.
Keywords:	RNA Interference, Double RNAi, Genetic Screen, Conjugation, Reprogramming, Epigenetics, Animals, Caenorhabditis elegans
Source:	bioRxiv
Publisher:	Cold Spring Harbor Laboratory Press
Article Number:	2020.09.01.276972
Date:	1 September 2020
Official Publication:	https://doi.org/10.1101/2020.09.01.276972

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