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Parallel genetics of regulatory sequences in vivo

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Item Type:Preprint
Title:Parallel genetics of regulatory sequences in vivo
Creators Name:Froehlich, J. and Uyar, B. and Herzog, M. and Theil, K. and Glažar, P. and Akalin, A. and Rajewsky, N.
Abstract:Understanding how regulatory sequences control gene expression is fundamental to explain how phenotypes arise in health and disease. Traditional reporter assays inform about function of individual regulatory elements, typically in isolation. However, regulatory elements must ultimately be understood by perturbing them within their genomic environment and developmental- or tissue-specific contexts. This is technically challenging; therefore, few regulatory elements have been characterized in vivo. Here, we used inducible Cas9 and multiplexed guide RNAs to create hundreds of mutations in enhancers/promoters and 3′ UTRs of 16 genes in C. elegans. To quantify the consequences of mutations on expression, we developed a targeted RNA sequencing strategy across hundreds of mutant animals. We were also able to systematically and quantitatively assign fitness cost to mutations. Finally, we identified and characterized sequence elements that strongly regulate phenotypic traits. Our approach enables highly parallelized, functional analysis of regulatory sequences in vivo.
Keywords:Animals, Caenorhabditis elegans
Source:bioRxiv
Publisher:Cold Spring Harbor Laboratory Press
Article Number:2020.07.28.224998
Date:29 July 2020
Official Publication:https://doi.org/

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