In situ functional dissection of RNA cis-regulatory elements by multiplex CRISPR-Cas9 genome engineering View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2017-12

AUTHORS

Qianxin Wu, Quentin R. V. Ferry, Toni A. Baeumler, Yale S. Michaels, Dimitrios M. Vitsios, Omer Habib, Roland Arnold, Xiaowei Jiang, Stefano Maio, Bruno R. Steinkraus, Marta Tapia, Paolo Piazza, Ni Xu, Georg A. Holländer, Thomas A. Milne, Jin-Soo Kim, Anton J. Enright, Andrew R. Bassett, Tudor A. Fulga

ABSTRACT

RNA regulatory elements (RREs) are an important yet relatively under-explored facet of gene regulation. Deciphering the prevalence and functional impact of this post-transcriptional control layer requires technologies for disrupting RREs without perturbing cellular homeostasis. Here we describe genome-engineering based evaluation of RNA regulatory element activity (GenERA), a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 platform for in situ high-content functional analysis of RREs. We use GenERA to survey the entire regulatory landscape of a 3'UTR, and apply it in a multiplex fashion to analyse combinatorial interactions between sets of miRNA response elements (MREs), providing strong evidence for cooperative activity. We also employ this technology to probe the functionality of an entire MRE network under cellular homeostasis, and show that high-resolution analysis of the GenERA dataset can be used to extract functional features of MREs. This study provides a genome editing-based multiplex strategy for direct functional interrogation of RNA cis-regulatory elements in a native cellular environment. More... »

PAGES

2109

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/s41467-017-00686-2

    DOI

    http://dx.doi.org/10.1038/s41467-017-00686-2

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1099597668

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/29235467


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