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

Mustapha Aouida; Ayman Eid; Magdy M. Mahfouz

doi:10.1016/j.biopen.2016.02.001

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

Mustapha Aouida, Ayman Eid, Magdy M. Mahfouz

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

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.

Original language	English (US)
Pages (from-to)	72-75
Number of pages	4
Journal	Biochimie Open
Volume	3
DOIs	https://doi.org/10.1016/j.biopen.2016.02.001
State	Published - Feb 25 2016

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: 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).

ASJC Scopus subject areas

Biochemistry

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10.1016/j.biopen.2016.02.001

https://repository.kaust.edu.sa/bitstream/10754/600212/1/1-s2.0-S2214008516000043-main.pdf

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title = "CRISPR/Cas9-mediated target validation of the Splicing Inhibitor Pladienolide B",

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.",

author = "Mustapha Aouida and Ayman Eid and Mahfouz, {Magdy M.}",

note = "KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: 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).",

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doi = "10.1016/j.biopen.2016.02.001",

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AU - Eid, Ayman

AU - Mahfouz, Magdy M.

N1 - KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: 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).

PY - 2016/2/25

Y1 - 2016/2/25

N2 - 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.

AB - 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.

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ER -

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

Abstract

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ASJC Scopus subject areas

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