Abstract

The Genetic Modification Core will generate genetically engineered human ES (hES) cell lines required for the four primary projects and the pilot projects of the Program. These include gene null, conditional null, and knock-in hES cell lines. In addition, a very efficient recombination medicated cassette exchange method will be used to generate gene overexpression and/or knockdown (by RNAi) hES cell lines. Gene constructs generated by the Program investigators will be electroporated into NIH-approved human ES cells (H1 or H9) and cultured with appropriate drugs to select for DNA transformants. The resulting hES cell colonies will be picked and expanded for genotype analysis. Genotype analysis will be performed by the Project investigators. After the genetically engineered hES cell lines are identified and confirmed they will be transferred to Core B for karyotype analysis, differentiation potential (if required) in vitro and in vivo by teratoma formation, and expansion for cell banking. All of the four primary projects propose to generate genetically modified hES cell lines. Thus, the Genetic Modification Core is an essential component of the Program.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM081627-05
Application #
8327220
Study Section
Special Emphasis Panel (ZGM1)
Project Start
Project End
2014-08-31
Budget Start
2011-09-01
Budget End
2014-08-31
Support Year
5
Fiscal Year
2011
Total Cost
$346,143
Indirect Cost

  Institution

Name
Baylor College of Medicine
Department
Type
DUNS #
051113330
City
Houston
State
TX
Country
United States
Zip Code
77030

  Publications

Fujita, Jun; Freire, Pablo; Coarfa, Cristian et al. (2017) Ronin Governs Early Heart Development by Controlling Core Gene Expression Programs. Cell Rep 21:1562-1573
Jain, Abhinav K; Xi, Yuanxin; McCarthy, Ryan et al. (2016) LncPRESS1 Is a p53-Regulated LncRNA that Safeguards Pluripotency by Disrupting SIRT6-Mediated De-acetylation of Histone H3K56. Mol Cell 64:967-981
Wang, Hongran; Wang, Xiaohong; Xu, Xueping et al. (2016) Germ Cell Nuclear Factor (GCNF) Represses Oct4 Expression and Globally Modulates Gene Expression in Human Embryonic Stem (hES) Cells. J Biol Chem 291:8644-52
Wang, Hongran; Xi, Yutao; Zheng, Yi et al. (2016) Generation of electrophysiologically functional cardiomyocytes from mouse induced pluripotent stem cells. Stem Cell Res 16:522-30
Skinner, Samuel O; Xu, Heng; Nagarkar-Jaiswal, Sonal et al. (2016) Single-cell analysis of transcription kinetics across the cell cycle. Elife 5:e12175
Tang, Mengfan; Li, Yujing; Zhang, Yi et al. (2015) Disease mutant analysis identifies a new function of DAXX in telomerase regulation and telomere maintenance. J Cell Sci 128:331-41
Lu, Weisi; Fang, Lekun; Ouyang, Bin et al. (2015) Actl6a protects embryonic stem cells from differentiating into primitive endoderm. Stem Cells 33:1782-93
Zheng, Ze-Yi; Tian, Lin; Bu, Wen et al. (2015) Wild-Type N-Ras, Overexpressed in Basal-like Breast Cancer, Promotes Tumor Formation by Inducing IL-8 Secretion via JAK2 Activation. Cell Rep 12:511-24
Sabour, Davood; Xu, Xueping; Chung, Arthur C K et al. (2014) Germ cell nuclear factor regulates gametogenesis in developing gonads. PLoS One 9:e103985
Li, Yujing; Fong, Ka-Wing; Tang, Mengfan et al. (2014) Fam118B, a newly identified component of Cajal bodies, is required for Cajal body formation, snRNP biogenesis and cell viability. J Cell Sci 127:2029-39

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