CRISPR/Cas9-mediated target validation of the Splicing Inhibitor Pladienolide B

Handle URI:
http://hdl.handle.net/10754/600212
Title:
CRISPR/Cas9-mediated target validation of the Splicing Inhibitor Pladienolide B
Authors:
Aouida, Mustapha ( 0000-0001-7945-5320 ) ; Eid, Ayman; Mahfouz, Magdy M. ( 0000-0002-0616-6365 )
Abstract:
CRISPR/Cas9 system confers molecular immunity in archeal and bacterial species against invading foreign nucleic acids. CRISPR/Cas9 system is used for genome engineering applications across diverse eukaryotic species. In this study, we demonstrate the utility of the CRISPR/Cas9 genome engineering system for drug target validation in human cells. Pladienolide B is a natural macrolide with antitumor activities mediated through the inhibition of pre-mRNA splicing. To validate the spliceosomal target of Pladienolide B, we employed the CRSIPR/Cas9 system to introduce targeted mutations in the subunits of the SF3B complex in the HEK293T cells. Our data reveal that targeted mutagenesis of the SF3b1 subunit exhibited higher levels of resistance to Pladienolide B. Therefore, our data validate the spliceosomal target of Pladienolide B and provide a proof of concept on using the CRISPR/Cas9 system for drug target identification and validation.
KAUST Department:
Laboratory for Genome Engineering; Biological Sciences; Center for Desert Agriculture
Citation:
CRISPR/Cas9-mediated target validation of the Splicing Inhibitor Pladienolide B 2016 Biochimie Open
Publisher:
Elsevier BV
Journal:
Biochimie Open
Issue Date:
24-Feb-2016
DOI:
10.1016/j.biopen.2016.02.001
Type:
Article
ISSN:
22140085
Sponsors:
We thank the Bioscience Core Facility at King Abdullah University of Science and Technology (KAUST) for technical assistance. We also thank the members of the genome engineering group at KAUST for their helpful discussions and technical assistance throughout the preparation of the manuscript. This research was funded King Abdullah University of Science and Technology (KAUST).
Additional Links:
http://linkinghub.elsevier.com/retrieve/pii/S2214008516000043
Appears in Collections:
Articles; Desert Agriculture Initiative

Full metadata record

DC FieldValue Language
dc.contributor.authorAouida, Mustaphaen
dc.contributor.authorEid, Aymanen
dc.contributor.authorMahfouz, Magdy M.en
dc.date.accessioned2016-02-28T13:09:00Zen
dc.date.available2016-02-28T13:09:00Zen
dc.date.issued2016-02-24en
dc.identifier.citationCRISPR/Cas9-mediated target validation of the Splicing Inhibitor Pladienolide B 2016 Biochimie Openen
dc.identifier.issn22140085en
dc.identifier.doi10.1016/j.biopen.2016.02.001en
dc.identifier.urihttp://hdl.handle.net/10754/600212en
dc.description.abstractCRISPR/Cas9 system confers molecular immunity in archeal and bacterial species against invading foreign nucleic acids. CRISPR/Cas9 system is used for genome engineering applications across diverse eukaryotic species. In this study, we demonstrate the utility of the CRISPR/Cas9 genome engineering system for drug target validation in human cells. Pladienolide B is a natural macrolide with antitumor activities mediated through the inhibition of pre-mRNA splicing. To validate the spliceosomal target of Pladienolide B, we employed the CRSIPR/Cas9 system to introduce targeted mutations in the subunits of the SF3B complex in the HEK293T cells. Our data reveal that targeted mutagenesis of the SF3b1 subunit exhibited higher levels of resistance to Pladienolide B. Therefore, our data validate the spliceosomal target of Pladienolide B and provide a proof of concept on using the CRISPR/Cas9 system for drug target identification and validation.en
dc.description.sponsorshipWe thank the Bioscience Core Facility at King Abdullah University of Science and Technology (KAUST) for technical assistance. We also thank the members of the genome engineering group at KAUST for their helpful discussions and technical assistance throughout the preparation of the manuscript. This research was funded King Abdullah University of Science and Technology (KAUST).en
dc.language.isoenen
dc.publisherElsevier BVen
dc.relation.urlhttp://linkinghub.elsevier.com/retrieve/pii/S2214008516000043en
dc.rightsArchived with thanks to Biochimie Open, Under a Creative Commons license, http://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjectCRIPSR/Cas9en
dc.subjectPladienolide Ben
dc.subjectspliceosomeen
dc.subjectpre-mRNA splicingen
dc.subjectdrug target validationen
dc.subjectdrug discoveryen
dc.titleCRISPR/Cas9-mediated target validation of the Splicing Inhibitor Pladienolide Ben
dc.typeArticleen
dc.contributor.departmentLaboratory for Genome Engineeringen
dc.contributor.departmentBiological Sciencesen
dc.contributor.departmentCenter for Desert Agricultureen
dc.identifier.journalBiochimie Openen
dc.eprint.versionPost-printen
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)en
kaust.authorAouida, Mustaphaen
kaust.authorEid, Aymanen
kaust.authorMahfouz, Magdy M.en
All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.