Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.

Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.

CRISPR/Cas9 is an enabling RNA-guided technology for genome targeting and engineering. An acute DNA binding constraint of the Cas9 protein is the Protospacer Adjacent Motif (PAM). Here we demonstrate that the PAM requirement can be exploited to specifically target single-nucleotide heterozygous muta...

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Main Authors: Yi Li, Saurabh Mendiratta, Kristina Ehrhardt, Neha Kashyap, Michael A White, Leonidas Bleris
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2016-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC4720446?pdf=render
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spelling doaj-282a024e7a274fce9ba6f8efbfac361f2020-11-25T02:31:45ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01111e014497010.1371/journal.pone.0144970Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.Yi LiSaurabh MendirattaKristina EhrhardtNeha KashyapMichael A WhiteLeonidas BlerisCRISPR/Cas9 is an enabling RNA-guided technology for genome targeting and engineering. An acute DNA binding constraint of the Cas9 protein is the Protospacer Adjacent Motif (PAM). Here we demonstrate that the PAM requirement can be exploited to specifically target single-nucleotide heterozygous mutations while exerting no aberrant effects on the wild-type alleles. Specifically, we target the heterozygous G13A activating mutation of KRAS in colorectal cancer cells and we show reversal of drug resistance to a MEK small-molecule inhibitor. Our study introduces a new paradigm in genome editing and therapeutic targeting via the use of gRNA to guide Cas9 to a desired protospacer adjacent motif.http://europepmc.org/articles/PMC4720446?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Yi Li
Saurabh Mendiratta
Kristina Ehrhardt
Neha Kashyap
Michael A White
Leonidas Bleris
spellingShingle Yi Li
Saurabh Mendiratta
Kristina Ehrhardt
Neha Kashyap
Michael A White
Leonidas Bleris
Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.
PLoS ONE
author_facet Yi Li
Saurabh Mendiratta
Kristina Ehrhardt
Neha Kashyap
Michael A White
Leonidas Bleris
author_sort Yi Li
title Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.
title_short Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.
title_full Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.
title_fullStr Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.
title_full_unstemmed Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions.
title_sort exploiting the crispr/cas9 pam constraint for single-nucleotide resolution interventions.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2016-01-01
description CRISPR/Cas9 is an enabling RNA-guided technology for genome targeting and engineering. An acute DNA binding constraint of the Cas9 protein is the Protospacer Adjacent Motif (PAM). Here we demonstrate that the PAM requirement can be exploited to specifically target single-nucleotide heterozygous mutations while exerting no aberrant effects on the wild-type alleles. Specifically, we target the heterozygous G13A activating mutation of KRAS in colorectal cancer cells and we show reversal of drug resistance to a MEK small-molecule inhibitor. Our study introduces a new paradigm in genome editing and therapeutic targeting via the use of gRNA to guide Cas9 to a desired protospacer adjacent motif.
url http://europepmc.org/articles/PMC4720446?pdf=render
work_keys_str_mv AT yili exploitingthecrisprcas9pamconstraintforsinglenucleotideresolutioninterventions
AT saurabhmendiratta exploitingthecrisprcas9pamconstraintforsinglenucleotideresolutioninterventions
AT kristinaehrhardt exploitingthecrisprcas9pamconstraintforsinglenucleotideresolutioninterventions
AT nehakashyap exploitingthecrisprcas9pamconstraintforsinglenucleotideresolutioninterventions
AT michaelawhite exploitingthecrisprcas9pamconstraintforsinglenucleotideresolutioninterventions
AT leonidasbleris exploitingthecrisprcas9pamconstraintforsinglenucleotideresolutioninterventions
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