Optimized base editors enable efficient editing in cells, organoids and mice

被引：259

作者：

Zafra, Maria Paz ^{[1
]}

Schatoff, Emma M. ^{[1
,2
]}

Katti, Alyna ^{[1
,3
]}

Foronda, Miguel ^{[1
]}

Breinig, Marco ^{[4
,5
]}

Schweitzer, Anabel Y. ^{[4
,5
]}

Simon, Amber ^{[1
]}

Han, Teng ^{[1
,3
]}

Goswami, Sukanya ^{[1
]}

Montgomery, Emma ^{[1
]}

Thibado, Jordana ^{[3
]}

Kastenhuber, Edward R. ^{[6
,7
]}

Sanchez-Rivera, Francisco J. ^{[6
]}

Shi, Junwei ^{[8
,9
]}

Vakoc, Christopher R. ^{[8
]}

Lowe, Scott W. ^{[6
,10
]}

Tschaharganeh, Darjus F. ^{[4
,5
]}

Dow, Lukas E. ^{[1
,3
,11
]}

机构：

[1] Weill Cornell Med, Dept Med, Sandra & Edward Meyer Canc Ctr, New York, NY 10065 USA

[2] Weill Cornell Rockefeller Sloan Kettering Tri Ins, New York, NY USA

[3] Weill Cornell Med, Weill Cornell Grad Sch Med Sci, New York, NY 10065 USA

[4] Univ Hosp, German Canc Res Ctr, DKFZ, Helmholtz Univ Grp Cell Plast & Epigenet Remod, Heidelberg, Germany

[5] Univ Hosp, Inst Pathol, Heidelberg, Germany

[6] Mem Sloan Kettering Canc Ctr, Cancer Biol & Genet, 1275 York Ave, New York, NY 10021 USA

[7] Gerstner Sloan Kettering Grad Sch Biomed Sci, New York, NY USA

[8] Cold Spring Harbor Lab, New York, NY USA

[9] Univ Penn, Perelman Sch Med, Dept Canc Biol, Philadelphia, PA 19104 USA

[10] Mem Sloan Kettering Canc Ctr, Howard Hughes Med Inst, 1275 York Ave, New York, NY 10021 USA

[11] Weill Cornell Med, Dept Biochem, New York, NY 10065 USA

来源：

NATURE BIOTECHNOLOGY | 2018年 / 36卷 / 09期

关键词：

CRISPR-CAS9; NUCLEASES; GENOMIC DNA; GENERATION; LIVER;

D O I：

10.1038/nbt.4194

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

CRISPR base editing enables the creation of targeted single-base conversions without generating double-stranded breaks. However, the efficiency of current base editors is very low in many cell types. We reengineered the sequences of BE3, BE4Gam, and xBE3 by codon optimization and incorporation of additional nuclear-localization sequences. Our collection of optimized constitutive and inducible base-editing vector systems dramatically improves the efficiency by which single-nucleotide variants can be created. The reengineered base editors enable target modification in a wide range of mouse and human cell lines, and intestinal organoids. We also show that the optimized base editors mediate efficient in vivo somatic editing in the liver in adult mice.

引用

页码：888 / +

页数：9

共 32 条

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