An Introduction to CRISPR Technology for Genome Activation and Repression in Mammalian Cells

Dan Du; Lei S Qi

doi:10.1101/pdb.top086835

An Introduction to CRISPR Technology for Genome Activation and Repression in Mammalian Cells

Cold Spring Harb Protoc. 2016 Jan 4;2016(1):pdb.top086835. doi: 10.1101/pdb.top086835.

Authors

Dan Du¹, Lei S Qi²

Affiliations

¹ Department of Bioengineering, Stanford University, Stanford, California 94305;
² Department of Bioengineering, Stanford University, Stanford, California 94305; Department of Chemical and Systems Biology, Stanford University, Stanford, California 94305; ChEM-H; Stanford University, Stanford, California 94305.

PMID: 26729914
DOI: 10.1101/pdb.top086835

Abstract

CRISPR interference/activation (CRISPRi/a) technology provides a simple and efficient approach for targeted repression or activation of gene expression in the mammalian genome. It is highly flexible and programmable, using an RNA-guided nuclease-deficient Cas9 (dCas9) protein fused with transcriptional regulators for targeting specific genes to effect their regulation. Multiple studies have shown how this method is an effective way to achieve efficient and specific transcriptional repression or activation of single or multiple genes. Sustained transcriptional modulation can be obtained by stable expression of CRISPR components, which enables directed reprogramming of cell fate. Here, we introduce the basics of CRISPRi/a technology for genome repression or activation.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Clustered Regularly Interspaced Short Palindromic Repeats*
Gene Expression Regulation*
Gene Targeting / methods*
Genome / genetics*
Humans
Mammals
RNA Interference / physiology*
RNA, Untranslated / genetics
RNA, Untranslated / metabolism

Substances

RNA, Untranslated

Grants and funding

OD017887/OD/NIH HHS/United States