Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), with their abilities to replicate indefinitely and maintain pluripotency, offer tremendous opportunities for the development of novel regenerative medicine-based therapies. iPSC technology, in particular, may provide the means to develop personalized therapeutic strategies in which a patient's own cells might be modified and used to treat their disease without the need for immunosuppression. Patient-derived ESCs and iPSCs will also provide critically important reagents for modeling and studying the biology of various diseases. Despite tremendous recent progress, many significant challenges remain to be addressed before ESCs and iPSCs can be used routinely for therapeutics. For example, methods for generating iPSCs are currently inefficient with considerable room for improvement. In addition, homologous recombination-mediated gene targeting for creating specific mutations works with low efficiency and is difficult to perform, particularly in human ESCs and iPSCs. Lastly, methods for robustly differentiating ESCs and iPSCs into cell types of interest are typically inefficient and yield cells that fail to exhibit all desired characteristics. The goal of this proposal is to employ engineered zinc finger technology to address these major challenges impeding research and therapeutic applications of ESCs and iPSCs. Specifically, this proposal will utilize the PI's expertise in engineered zinc finger technology to address three important challenges for stem cell biology and research: (1) developing more efficient methods for iPSC generation, (2) enabling high efficiency ZFN-induced gene targeting in human ESCs and iPSCs, and (3) creating more efficient methods for differentiating stem cells into desired cell types of interest. Public Health Relevance: This application seeks to address significant challenges that currently limit research and therapeutic applications of human embryonic stem cells a

National Institute of Health (NIH)
Office of The Director, National Institutes of Health (OD)
NIH Director’s Pioneer Award (NDPA) (DP1)
Project #
Application #
Study Section
Special Emphasis Panel (ZGM1-NDPA-B (01))
Program Officer
Wehrle, Janna P
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Massachusetts General Hospital
United States
Zip Code
Keung, Albert J; Joung, J Keith; Khalil, Ahmad S et al. (2015) Chromatin regulation at the frontier of synthetic biology. Nat Rev Genet 16:159-71
Spisák, Sándor; Lawrenson, Kate; Fu, Yanfang et al. (2015) CAUSEL: an epigenome- and genome-editing pipeline for establishing function of noncoding GWAS variants. Nat Med 21:1357-63
Kiskinis, Evangelos; Sandoe, Jackson; Williams, Luis A et al. (2014) Pathways disrupted in human ALS motor neurons identified through genetic correction of mutant SOD1. Cell Stem Cell 14:781-95
Osborn, Mark J; Starker, Colby G; McElroy, Amber N et al. (2013) TALEN-based gene correction for epidermolysis bullosa. Mol Ther 21:1151-9
Hwang, Woong Y; Fu, Yanfang; Reyon, Deepak et al. (2013) Heritable and precise zebrafish genome editing using a CRISPR-Cas system. PLoS One 8:e68708
Reyon, Deepak; Maeder, Morgan L; Khayter, Cyd et al. (2013) Engineering customized TALE nucleases (TALENs) and TALE transcription factors by fast ligation-based automatable solid-phase high-throughput (FLASH) assembly. Curr Protoc Mol Biol Chapter 12:Unit 12.16
Maeder, Morgan L; Linder, Samantha J; Reyon, Deepak et al. (2013) Robust, synergistic regulation of human gene expression using TALE activators. Nat Methods 10:243-5
Reyon, Deepak; Khayter, Cyd; Regan, Maureen R et al. (2012) Engineering designer transcription activator-like effector nucleases (TALENs) by REAL or REAL-Fast assembly. Curr Protoc Mol Biol Chapter 12:Unit 12.15
Cade, Lindsay; Reyon, Deepak; Hwang, Woong Y et al. (2012) Highly efficient generation of heritable zebrafish gene mutations using homo- and heterodimeric TALENs. Nucleic Acids Res 40:8001-10
Moore, Finola E; Reyon, Deepak; Sander, Jeffry D et al. (2012) Improved somatic mutagenesis in zebrafish using transcription activator-like effector nucleases (TALENs). PLoS One 7:e37877

Showing the most recent 10 out of 35 publications