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...

Descripción completa

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
Descripción
Sumario: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.

Ejemplares similares