Tonge, DP ORCID: https://orcid.org/0000-0002-3499-2752, Darling, D, Farzaneh, F and Williams, GT (2022) Whole-genome-scale identification of novel non-protein-coding RNAs controlling cell proliferation and survival through a functional forward genetics strategy. Scientific Reports, 12 (1).

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Abstract

<jats:title>Abstract</jats:title><jats:p>Identification of cell fate-controlling lncRNAs is essential to our understanding of molecular cell biology. Here we present a human genome-scale forward-genetics approach for the identification of lncRNAs based on gene function. This approach can identify genes that play a causal role, and immediately distinguish them from those that are differentially expressed but do not affect cell function. Our genome-scale library plus next-generation-sequencing and bioinformatic approach, radically upscales the breadth and rate of functional ncRNA discovery. Human gDNA was digested to produce a lentiviral expression library containing inserts in both sense and anti-sense orientation. The library was used to transduce human Jurkat T-leukaemic cells. Cell populations were selected using continuous culture ± anti-FAS IgM, and sequencing used to identify sequences controlling cell proliferation. This strategy resulted in the identification of thousands of new sequences based solely on their function including many ncRNAs previously identified as being able to modulate cell survival or to act as key cancer regulators such as <jats:italic>AC084816.1</jats:italic>*, <jats:italic>AC097103.2</jats:italic>, <jats:italic>AC087473.1</jats:italic>, <jats:italic>CASC15*</jats:italic>, <jats:italic>DLEU1</jats:italic>*, <jats:italic>ENTPD1-AS1</jats:italic>*, <jats:italic>HULC</jats:italic>*, <jats:italic>MIRLET7BHG</jats:italic>*, <jats:italic>PCAT-1</jats:italic>, <jats:italic>SChLAP1</jats:italic>, and <jats:italic>TP53TG1</jats:italic>. Independent validation confirmed 4 out of 5 sequences that were identified by this strategy, conferred a striking resistance to anti-FAS IgM-induced apoptosis.</jats:p>

Item Type: Article
Additional Information: Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Subjects: Q Science > QD Chemistry > QD415 Biochemistry
R Medicine > R Medicine (General)
R Medicine > R Medicine (General) > R735 Medical education. Medical schools. Research
R Medicine > RS Pharmacy and materia medica
Depositing User: Symplectic
Date Deposited: 13 Jan 2022 10:07
Last Modified: 13 Jan 2022 10:07
URI: https://eprints.keele.ac.uk/id/eprint/10497

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