A TALEN Genome-Editing System for Generating Human Stem Cell-Based Disease Models

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• 作者：Qiurong Ding¹ ; ¹⁰ ; Youn-Kyoung Lee¹ ; ¹⁰ ; Esperance A.K. Schaefer³ ; ⁵ ; ¹⁰ ; Derek T. Peters¹ ; ⁵ ; Adrian Veres¹ ; ⁵ ; Kevin Kim¹ ; Nicolas Kuperwasser¹ ; ⁶ ; Daniel L. Motola³ ; ⁵ ; Torsten B. Meissner¹ ; William T. Hendriks¹ ; ² ; Marta Trevisan¹ ; ² ; Rajat M. Gupta¹ ; ⁵ ; ⁶ ; Annie Moisan⁷ ; Eric Banks⁸ ; Max Friesen¹ ; Robert T. Schinzel¹ ; Fang Xia¹ ; Alexander Tang¹ ; Yulei Xia¹ ; Emmanuel Figueroa¹ ; Amy Wann¹ ; Tim Ahfeldt¹ ; Laurence Daheron¹ ; ² ; Feng Zhang⁸ ; ⁹ ; Lee L. Rubin¹ ; ² ; Lee F. Peng³ ; ⁵ ; Raymond T. Chung³ ; ⁵ ; Kiran Musunuru¹ ; ² ; ⁵ ; ⁶ ; ⁸ ; ¹¹ ; ^{kiranmusunuru@gmail.com} ; Chad A. Cowan¹ ; ² ; ⁴ ; ⁵ ; ⁸ ; ¹¹ ; ^{ccowan@fas.harvard.edu}
• 刊名：Cell Stem Cell
• 出版年：2013
• 出版时间：7 February, 2013
• 年：2013
• 卷：12
• 期：2
• 页码：238-251
• 全文大小：2171 K

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Summary

Transcription activator-like effector nucleases (TALENs) are a new class of engineered nucleases that are easier to design to cleave at desired sites in a genome than previous types of nucleases. We report here the use of TALENs to rapidly and efficiently generate mutant alleles of 15 genes in cultured somatic cells or human pluripotent stem cells, the latter for which we differentiated both the targeted lines and isogenic control lines into various metabolic cell types. We demonstrate cell-autonomous phenotypes directly linked to disease¡ªdyslipidemia, insulin resistance, hypoglycemia, lipodystrophy, motor-neuron death, and hepatitis C infection. We found little evidence of TALEN off-target effects, but each clonal line nevertheless harbors a significant number of unique mutations. Given the speed and ease with which we were able to derive and characterize these cell lines, we anticipate TALEN-mediated genome editing of human cells becoming a mainstay for the investigation of human biology and disease.