title: A TALEN genome-editing system for generating human stem cell-based disease models.
abstract: 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-dyslipidemia, 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.
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA.
authors.forename: Qiurong
authors.initials: Q
authors.lastname: Ding
authors.forename: Youn-Kyoung
authors.initials: YK
authors.lastname: Lee
authors.forename: Esperance A K
authors.initials: EA
authors.lastname: Schaefer
authors.forename: Derek T
authors.initials: DT
authors.lastname: Peters
authors.forename: Adrian
authors.initials: A
authors.lastname: Veres
authors.forename: Kevin
authors.initials: K
authors.lastname: Kim
authors.forename: Nicolas
authors.initials: N
authors.lastname: Kuperwasser
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Motola
authors.forename: Torsten B
authors.initials: TB
authors.lastname: Meissner
authors.forename: William T
authors.initials: WT
authors.lastname: Hendriks
authors.forename: Marta
authors.initials: M
authors.lastname: Trevisan
authors.forename: Rajat M
authors.initials: RM
authors.lastname: Gupta
authors.forename: Annie
authors.initials: A
authors.lastname: Moisan
authors.forename: Eric
authors.initials: E
authors.lastname: Banks
authors.forename: Max
authors.initials: M
authors.lastname: Friesen
authors.forename: Robert T
authors.initials: RT
authors.lastname: Schinzel
authors.forename: Fang
authors.initials: F
authors.lastname: Xia
authors.forename: Alexander
authors.initials: A
authors.lastname: Tang
authors.forename: Yulei
authors.initials: Y
authors.lastname: Xia
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Figueroa
authors.forename: Amy
authors.initials: A
authors.lastname: Wann
authors.forename: Tim
authors.initials: T
authors.lastname: Ahfeldt
authors.forename: Laurence
authors.initials: L
authors.lastname: Daheron
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Lee L
authors.initials: LL
authors.lastname: Rubin
authors.forename: Lee F
authors.initials: LF
authors.lastname: Peng
authors.forename: Raymond T
authors.initials: RT
authors.lastname: Chung
authors.forename: Kiran
authors.initials: K
authors.lastname: Musunuru
authors.forename: Chad A
authors.initials: CA
authors.lastname: Cowan
mesh_terms: D000483:Alleles; D003851:Deoxyribonucleases / Q000235:genetics*; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D009154:Mutation; D013234:Stem Cells / Q000201:enzymology*
chemical_list: D003851:Deoxyribonucleases