Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease

We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct sgRNA constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14 15 bp target sequences and MS2 binding loops, can activate gene expression using an active Cas9 nuclease, withou...

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Detalles Bibliográficos
Autores principales: Dahlman, James E., Abudayyeh, Omar O., Joung, Julia, Gootenberg, Jonathan S., Zhang, Feng, Konermann, Silvana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4747789/
https://www.ncbi.nlm.nih.gov/pubmed/26436575
http://dx.doi.org/10.1038/nbt.3390
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author Dahlman, James E.
Abudayyeh, Omar O.
Joung, Julia
Gootenberg, Jonathan S.
Zhang, Feng
Konermann, Silvana
author_facet Dahlman, James E.
Abudayyeh, Omar O.
Joung, Julia
Gootenberg, Jonathan S.
Zhang, Feng
Konermann, Silvana
author_sort Dahlman, James E.
collection PubMed
description We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct sgRNA constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14 15 bp target sequences and MS2 binding loops, can activate gene expression using an active Cas9 nuclease, without inducing DSBs. We use these ‘dead RNAs’ to perform orthogonal gene knockout and transcriptional activation in human cells.
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spelling pubmed-47477892016-05-01 Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease Dahlman, James E. Abudayyeh, Omar O. Joung, Julia Gootenberg, Jonathan S. Zhang, Feng Konermann, Silvana Nat Biotechnol Article We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct sgRNA constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14 15 bp target sequences and MS2 binding loops, can activate gene expression using an active Cas9 nuclease, without inducing DSBs. We use these ‘dead RNAs’ to perform orthogonal gene knockout and transcriptional activation in human cells. 2015-11 /pmc/articles/PMC4747789/ /pubmed/26436575 http://dx.doi.org/10.1038/nbt.3390 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Dahlman, James E.
Abudayyeh, Omar O.
Joung, Julia
Gootenberg, Jonathan S.
Zhang, Feng
Konermann, Silvana
Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease
title Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease
title_full Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease
title_fullStr Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease
title_full_unstemmed Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease
title_short Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease
title_sort orthogonal gene knock out and activation with a catalytically active cas9 nuclease
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4747789/
https://www.ncbi.nlm.nih.gov/pubmed/26436575
http://dx.doi.org/10.1038/nbt.3390
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