Cas9 As a Versatile Tool for Engineering Biology

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2013 Oct 1

PMID 24076990

Citations 604

Authors

Prashant Mali

Kevin M Esvelt

George M Church

Affiliations

Soon will be listed here.

Abstract

RNA-guided Cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have dramatically transformed our ability to edit the genomes of diverse organisms. We believe tools and techniques based on Cas9, a single unifying factor capable of colocalizing RNA, DNA and protein, will grant unprecedented control over cellular organization, regulation and behavior. Here we describe the Cas9 targeting methodology, detail current and prospective engineering advances and suggest potential applications ranging from basic science to the clinic.

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References

Mali P, Aach J, Lee J, Levner D, Nip L, Church G . Barcoding cells using cell-surface programmable DNA-binding domains. Nat Methods. 2013; 10(5):403-6. PMC: 3641172. DOI: 10.1038/nmeth.2407. View

Maeder M, Linder S, Reyon D, Angstman J, Fu Y, Sander J . Robust, synergistic regulation of human gene expression using TALE activators. Nat Methods. 2013; 10(3):243-5. PMC: 3584229. DOI: 10.1038/nmeth.2366. View

Hsu P, Scott D, Weinstein J, Ran F, Konermann S, Agarwala V . DNA targeting specificity of RNA-guided Cas9 nucleases. Nat Biotechnol. 2013; 31(9):827-32. PMC: 3969858. DOI: 10.1038/nbt.2647. View

Rothemund P . Folding DNA to create nanoscale shapes and patterns. Nature. 2006; 440(7082):297-302. DOI: 10.1038/nature04586. View

Lanctot C, Cheutin T, Cremer M, Cavalli G, Cremer T . Dynamic genome architecture in the nuclear space: regulation of gene expression in three dimensions. Nat Rev Genet. 2007; 8(2):104-15. DOI: 10.1038/nrg2041. View

Esvelt K, Carlson J, Liu D . A system for the continuous directed evolution of biomolecules. Nature. 2011; 472(7344):499-503. PMC: 3084352. DOI: 10.1038/nature09929. View

Urnov F, Miller J, Lee Y, Beausejour C, Rock J, Augustus S . Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature. 2005; 435(7042):646-51. DOI: 10.1038/nature03556. View

Terns M, Terns R . CRISPR-based adaptive immune systems. Curr Opin Microbiol. 2011; 14(3):321-7. PMC: 3119747. DOI: 10.1016/j.mib.2011.03.005. View

Pattanayak V, Lin S, Guilinger J, Ma E, Doudna J, Liu D . High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity. Nat Biotechnol. 2013; 31(9):839-43. PMC: 3782611. DOI: 10.1038/nbt.2673. View

10.

Hwang W, Fu Y, Reyon D, Maeder M, Tsai S, Sander J . Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol. 2013; 31(3):227-9. PMC: 3686313. DOI: 10.1038/nbt.2501. View

11.

Maeder M, Linder S, Cascio V, Fu Y, Ho Q, Joung J . CRISPR RNA-guided activation of endogenous human genes. Nat Methods. 2013; 10(10):977-9. PMC: 3794058. DOI: 10.1038/nmeth.2598. View

12.

Perez-Pinera P, Kocak D, Vockley C, Adler A, Kabadi A, Polstein L . RNA-guided gene activation by CRISPR-Cas9-based transcription factors. Nat Methods. 2013; 10(10):973-6. PMC: 3911785. DOI: 10.1038/nmeth.2600. View

13.

Livet J, Weissman T, Kang H, Draft R, Lu J, Bennis R . Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system. Nature. 2007; 450(7166):56-62. DOI: 10.1038/nature06293. View

14.

Li W, Teng F, Li T, Zhou Q . Simultaneous generation and germline transmission of multiple gene mutations in rat using CRISPR-Cas systems. Nat Biotechnol. 2013; 31(8):684-6. DOI: 10.1038/nbt.2652. View

15.

Hale C, Majumdar S, Elmore J, Pfister N, Compton M, Olson S . Essential features and rational design of CRISPR RNAs that function with the Cas RAMP module complex to cleave RNAs. Mol Cell. 2012; 45(3):292-302. PMC: 3278580. DOI: 10.1016/j.molcel.2011.10.023. View

16.

Gratz S, Cummings A, Nguyen J, Hamm D, Donohue L, Harrison M . Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Genetics. 2013; 194(4):1029-35. PMC: 3730909. DOI: 10.1534/genetics.113.152710. View

17.

Jiang W, Bikard D, Cox D, Zhang F, Marraffini L . RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol. 2013; 31(3):233-9. PMC: 3748948. DOI: 10.1038/nbt.2508. View

18.

Fusco D, Accornero N, Lavoie B, Shenoy S, Blanchard J, Singer R . Single mRNA molecules demonstrate probabilistic movement in living mammalian cells. Curr Biol. 2003; 13(2):161-167. PMC: 4764064. DOI: 10.1016/s0960-9822(02)01436-7. View

19.

Verma I, Somia N . Gene therapy -- promises, problems and prospects. Nature. 1997; 389(6648):239-42. DOI: 10.1038/38410. View

20.

Wang H, Yang H, Shivalila C, Dawlaty M, Cheng A, Zhang F . One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell. 2013; 153(4):910-8. PMC: 3969854. DOI: 10.1016/j.cell.2013.04.025. View