Journal
NATURE BIOTECHNOLOGY
Volume 33, Issue 11, Pages 1159-U147Publisher
NATURE RESEARCH
DOI: 10.1038/nbt.3390
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Funding
- Life Science Research Foundation post-doctoral fellowship of the Cystic Fibrosis Foundation
- Friends of the McGovern Institute Fellowship
- Department of Energy (DOE) Computational Science Graduate Fellowship
- National Institute of Mental Health (NIMH) [1DP1-MH100706]
- David R. Cheng and Bob Metcalfe
- Poitras Foundation
- Vallee Foundation
- Simons Foundation
- Paul G. Allen Foundation
- New York Stem Cell Foundation
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We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing doublestranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells.
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