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Genome engineering using site-specific nucleases

日期: 2013-10-28
学术报告
题目:Genome engineering using site-specific nucleases
报告人:Haoyi Wang, Ph.D
Whitehead Institute for Biomedical Research
Massachusetts Institute of Technology
时间:2013年11月7日(周四)下午15:00 PM
地点:生命学院411室
Targeted genetic engineering of mice and human pluripotent stem cells is essential for understanding gene function in development and disease. Gene targeting of human pluripotent stem cells by homologous recombination is inefficient. Using Transcription activator-like effector nucleases (TALENs), we developed strategies to modify genome of human pluripotent cells, which allows for establishment of novel disease models and screen platforms. In addition, we produced the world first mice harboring targeted mutations and insertions in Y-linked genes, providing a novel and general approach for genetic manipulation of the Y chromosome.
Furthermore, using CRISPR/Cas system we achieved multiplexed genome editing in mES cells and mouse with high efficiency. This method allows the one step generation of animals carrying mutations in multiple genes, an approach that will greatly accelerate the in vivo study of functionally redundant genes and of epistatic gene interactions. Finally, we achieved one step generation of mice carrying endogenous reporters as well as the derivation of conditional mutant mice using CRISPR/Cas mediated genome editing.
In addition to genetic manipulations, we developed a modification of CRISPR technology to regulate endogenous gene expression epigenetically. We succeeded in simultaneous activation of multiple endogenous genes with defined stoichiometry. This opens up exciting opportunities for system biology, as it allows for the specific manipulation of transcriptional networks.
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