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  • Radical acceleration of nuclear reprogramming by chromatin remodeling with the transactivation domain of MyoD.

Radical acceleration of nuclear reprogramming by chromatin remodeling with the transactivation domain of MyoD.

Stem cells (Dayton, Ohio) (2011-07-07)
Hiroyuki Hirai, Tetsuya Tani, Nobuko Katoku-Kikyo, Steven Kellner, Peter Karian, Meri Firpo, Nobuaki Kikyo
ABSTRACT

Induced pluripotent stem cells (iPSCs) can be created by reprogramming differentiated cells through introduction of defined genes, most commonly Oct4, Sox2, Klf4, and c-Myc (OSKM). However, this process is slow and extremely inefficient. Here, we demonstrate radical acceleration of iPSC creation with a fusion gene between Oct4 and the powerful transactivation domain (TAD) of MyoD (M(3)O). Transduction of M(3) O as well as Sox2, Klf4, and c-Myc into fibroblasts effectively remodeled patterns of DNA methylation, chromatin accessibility, histone modifications, and protein binding at pluripotency genes, raising the efficiency of making mouse and human iPSCs more than 50-fold in comparison to OSKM. These results identified that one of the most critical barriers to iPSC creation is poor chromatin accessibility and protein recruitment to pluripotency genes. The MyoD TAD has a capability of overcoming this problem. Our approach of fusing TADs to unrelated transcription factors has far-reaching implications as a powerful tool for transcriptional reprogramming beyond application to iPSC technology.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Alkaline Phosphatase Detection Kit, This Alkaline Phosphatase Detection Kit is a specific & sensitive tool for the phenotypic assessment of Embryonic Stem (ES) cell differentiation by the determination of AP activity.
Sigma-Aldrich
EmbryoMax® Acidic Tyrode′s Solution, The EmbryoMax Acidic Tyrode′s Solution is available in a 50 mL format and has been optimized and validated for Embryo Culture.
Sigma-Aldrich
EmbryoMax® FHM Mouse Embryo Media, (1X), Liquid, w/o Phenol Red
Sigma-Aldrich
EZ-Magna ChIP® G - Chromatin Immunoprecipitation Kit, Single day chromatin immunoprecipitation (ChIP) kit containing all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions using magnetic G beads. Control primers included.
Sigma-Aldrich
EmbryoMax® KSOM Mouse Embryo Media, (1X), Liquid, with 1/2 Amino Acids & Phenol Red