Dataset Information

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.

SUBMITTER: Ding Q

PROVIDER: S-EPMC3570604 | biostudies-other | 2013 Feb

REPOSITORIES: biostudies-other

ACCESS DATA

Publications

A TALEN genome-editing system for generating human stem cell-based disease models.

Ding Qiurong Q Lee Youn-Kyoung YK Schaefer Esperance A K EA Peters Derek T DT Veres Adrian A Kim Kevin K Kuperwasser Nicolas N Motola Daniel L DL Meissner Torsten B TB Hendriks William T WT Trevisan Marta M Gupta Rajat M RM Moisan Annie A Banks Eric E Friesen Max M Schinzel Robert T RT Xia Fang F Tang Alexander A Xia Yulei Y Figueroa Emmanuel E Wann Amy A Ahfeldt Tim T Daheron Laurence L Zhang Feng F Rubin Lee L LL Peng Lee F LF Chung Raymond T RT Musunuru Kiran K Cowan Chad A CA

Cell stem cell 20121213 2

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-auto ...[more]

PMID: 23246482

Similar Datasets

Project description:RationaleTargeted genetic engineering using programmable nucleases such as transcription activator-like effector nucleases (TALENs) is a valuable tool for precise, site-specific genetic modification in the human genome.ObjectiveThe emergence of novel technologies such as human induced pluripotent stem cells (iPSCs) and nuclease-mediated genome editing represent a unique opportunity for studying cardiovascular diseases in vitro.Methods and resultsBy incorporating extensive literature and database searches, we designed a collection of TALEN constructs to knockout 88 human genes that are associated with cardiomyopathies and congenital heart diseases. The TALEN pairs were designed to induce double-strand DNA break near the starting codon of each gene that either disrupted the start codon or introduced a frameshift mutation in the early coding region, ensuring faithful gene knockout. We observed that all the constructs were active and disrupted the target locus at high frequencies. To illustrate the utility of the TALEN-mediated knockout technique, 6 individual genes (TNNT2, LMNA/C, TBX5, MYH7, ANKRD1, and NKX2.5) were knocked out with high efficiency and specificity in human iPSCs. By selectively targeting a pathogenic mutation (TNNT2 p.R173W) in patient-specific iPSC-derived cardiac myocytes, we demonstrated that the knockout strategy ameliorates the dilated cardiomyopathy phenotype in vitro. In addition, we modeled the Holt-Oram syndrome in iPSC-cardiac myocytes in vitro and uncovered novel pathways regulated by TBX5 in human cardiac myocyte development.ConclusionsCollectively, our study illustrates the powerful combination of iPSCs and genome editing technologies for understanding the biological function of genes, and the pathological significance of genetic variants in human cardiovascular diseases. The methods, strategies, constructs, and iPSC lines developed in this study provide a validated, readily available resource for cardiovascular research.

Dataset Information

A TALEN genome-editing system for generating human stem cell-based disease models.

Publications

A TALEN genome-editing system for generating human stem cell-based disease models.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets