PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing

In CRISPR-Cas9 genome editing, the underlying principles for selecting guide RNA (gRNA) sequences that would ensure for efficient target site modification remain poorly understood. Here we show that target sites harbouring multiple protospacer adjacent motifs (PAMs) are refractory to Cas9-mediated r...

Descripción completa

Detalles Bibliográficos
Autores principales: Malina, Abba, Cameron, Christopher J. F., Robert, Francis, Blanchette, Mathieu, Dostie, Josée, Pelletier, Jerry
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4686818/
https://www.ncbi.nlm.nih.gov/pubmed/26644285
http://dx.doi.org/10.1038/ncomms10124
_version_ 1782406505182527488
author Malina, Abba
Cameron, Christopher J. F.
Robert, Francis
Blanchette, Mathieu
Dostie, Josée
Pelletier, Jerry
author_facet Malina, Abba
Cameron, Christopher J. F.
Robert, Francis
Blanchette, Mathieu
Dostie, Josée
Pelletier, Jerry
author_sort Malina, Abba
collection PubMed
description In CRISPR-Cas9 genome editing, the underlying principles for selecting guide RNA (gRNA) sequences that would ensure for efficient target site modification remain poorly understood. Here we show that target sites harbouring multiple protospacer adjacent motifs (PAMs) are refractory to Cas9-mediated repair in situ. Thus we refine which substrates should be avoided in gRNA design, implicating PAM density as a novel sequence-specific feature that inhibits in vivo Cas9-driven DNA modification.
format Online
Article
Text
id pubmed-4686818
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-46868182016-01-07 PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing Malina, Abba Cameron, Christopher J. F. Robert, Francis Blanchette, Mathieu Dostie, Josée Pelletier, Jerry Nat Commun Article In CRISPR-Cas9 genome editing, the underlying principles for selecting guide RNA (gRNA) sequences that would ensure for efficient target site modification remain poorly understood. Here we show that target sites harbouring multiple protospacer adjacent motifs (PAMs) are refractory to Cas9-mediated repair in situ. Thus we refine which substrates should be avoided in gRNA design, implicating PAM density as a novel sequence-specific feature that inhibits in vivo Cas9-driven DNA modification. Nature Publishing Group 2015-12-08 /pmc/articles/PMC4686818/ /pubmed/26644285 http://dx.doi.org/10.1038/ncomms10124 Text en Copyright © 2015, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Malina, Abba
Cameron, Christopher J. F.
Robert, Francis
Blanchette, Mathieu
Dostie, Josée
Pelletier, Jerry
PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing
title PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing
title_full PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing
title_fullStr PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing
title_full_unstemmed PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing
title_short PAM multiplicity marks genomic target sites as inhibitory to CRISPR-Cas9 editing
title_sort pam multiplicity marks genomic target sites as inhibitory to crispr-cas9 editing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4686818/
https://www.ncbi.nlm.nih.gov/pubmed/26644285
http://dx.doi.org/10.1038/ncomms10124
work_keys_str_mv AT malinaabba pammultiplicitymarksgenomictargetsitesasinhibitorytocrisprcas9editing
AT cameronchristopherjf pammultiplicitymarksgenomictargetsitesasinhibitorytocrisprcas9editing
AT robertfrancis pammultiplicitymarksgenomictargetsitesasinhibitorytocrisprcas9editing
AT blanchettemathieu pammultiplicitymarksgenomictargetsitesasinhibitorytocrisprcas9editing
AT dostiejosee pammultiplicitymarksgenomictargetsitesasinhibitorytocrisprcas9editing
AT pelletierjerry pammultiplicitymarksgenomictargetsitesasinhibitorytocrisprcas9editing

Ejemplares similares