Unraveling the potential of CRISPR-Cas9 for gene therapy

Abstract

The molecular machinery from the prokaryotic clustered regularly interspaced short palindromic repeats (CRISPR)-Cas immune system has broadly been repurposed for genome editing in eukaryotes. In particular, the sequence-specific Cas9 endonuclease can be flexibly harnessed for the genesis of precise double-stranded DNA breaks, using single guide RNAs that are readily programmable. The endogenous DNA repair machinery subsequently generates genome modifications, either by random insertion or deletions using non-homologous end joining (NHEJ), or designed integration of mutations or genetic material using homology-directed repair (HDR) templates. This technology has opened new avenues for the investigation of genetic diseases in general, and for gene therapy applications in particular.

Keywords:

• Cas9
• CRISPR
• gene therapy
• genome editing

Acknowledgements

R Barrangou is supported by start up funds from North Carolina State University. The authors would like to thank their many colleagues and collaborators in the CRISPR field for their insights into these fantastic molecular systems. The authors would also apologetically like to point out that it has become increasingly challenging to cite all desirable CRISPR literature, and that the reference limit herein has compounded this issue.

Declaration of interest

R Barrangou is an inventor on patents related to the use of CRISPR, is a member of the board of directors of Caribou Biosciences and is co-founder of Intellia Therapeutics. AP May is an inventor on patents related to the use of CRISPR, holds office in Caribou Biosciences and is co-founder of Intellia Therapeutics. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

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