2014-Present Investigator, Institute of Neuroscience, CAS
2012 – 2014 Postdoctoral Associate, Laboratory of Dr. Rudolf Jaenisch, Whitehead Institute, MIT, Cambridge, MA, USA
2007 – 2012 Ph.D. candidate, Laboratory of Dr. Jinsong Li, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China2003 – 2007 B.S., School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China
|1.Yang, H.*, Wang, H.*, Shivalila, C.S.*, Cheng, A.W., Shi, L., and Jaenisch, R. (2013). One-Step Generation of Mice Carrying Reporter and Conditional Alleles by CRISPR/Cas-Mediated Genome Engineering. Cell 154, 1370-1379.|
|2.Cheng, A.W., Wang, H., Yang, H., Shi, L., Katz, Y., Theunissen, T.W., Rangarajan, S., Shivalila, C.S., Dadon, D.B., and Jaenisch, R. (2013). Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system. Cell research 23, 1163-1171.|
|3.Wang, H.*, Yang, H.*, Shivalila, C.S.*, Dawlaty, M.M., Cheng, A.W., Zhang, F., and Jaenisch, R. (2013). One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153, 910-918.|
|4.Yang, H.*, Liu, Z.*, Ma, Y.*, Zhong, C., Yin, Q., Zhou, C., Shi, L., Cai, Y., Zhao, H., Wang, H., et al. (2013). Generation of haploid embryonic stem cells from Macaca fascicularis monkey parthenotes. Cell research 23, 1187-1200.|
|5.Jiang, J., Lv, W., Ye, X., Wang, L., Zhang, M., Yang, H., Okuka, M., Zhou, C., Zhang, X., Liu, L., et al. (2013). Zscan4 promotes genomic stability during reprogramming and dramatically improves the quality of iPS cells as demonstrated by tetraploid complementation. Cell research 23, 92-106.|
|6.Shi, L.*, Yang, H.*, and Li, J. (2012). Haploid embryonic stem cells: an ideal tool for mammalian genetic analyses. Protein & cell 3, 806-810. (Review)|
|7.Yang, H.*, Shi, L.*, Wang, B.A.*, Liang, D., Zhong, C., Liu, W., Nie, Y., Liu, J., Zhao, J., Gao, X., et al. (2012). Generation of genetically modified mice by oocyte injection of androgenetic haploid embryonic stem cells. Cell 149, 605-617.|
|8.Gu, T.P.*, Guo, F.*, Yang, H.*, Wu, H.P., Xu, G.F., Liu, W., Xie, Z.G., Shi, L., He, X., Jin, S.G., et al. (2011). The role of Tet3 DNA dioxygenase in epigenetic reprogramming by oocytes. Nature 477, 606-610.|
|9.Jiang, J., Ding, G., Lin, J., Zhang, M., Shi, L., Lv, W., Yang, H., Xiao, H., Pei, G., Li, Y., et al. (2011). Different developmental potential of pluripotent stem cells generated by different reprogramming strategies. Journal of molecular cell biology 3, 197-199.|
|10.Yang, H.*, Shi, L.*, Chen, C.D., and Li, J. (2011). Mice generated after round spermatid injection into haploid two-cell blastomeres. Cell Res 21, 854-858.|
|11.Lin, J., Shi, L., Zhang, M., Yang, H., Qin, Y., Zhang, J., Gong, D., Zhang, X., Li, D., and Li, J. (2011). Defects in trophoblast cell lineage account for the impaired in vivo development of cloned embryos generated by somatic nuclear transfer. Cell Stem Cell 8, 371-375.|
|12.Yang, H., Shi, L., Zhang, S., Ling, J., Jiang, J., and Li, J. (2010). High-efficiency somatic reprogramming induced by intact MII oocytes. Cell Res 20, 1034-1042.|
Genetically modified animals represent a crucial tool for understanding gene function in development and disease. In conventional gene-targeting methods to generate mutant animals, mutations are introduced through homologous recombination in embryonic stem (ES) cells. Targeted ES cells injected into wild-type (WT) blastocysts can contribute to the germline of chimeric animals, generating animals containing the targeted gene modification. It is costly and time consuming to produce single-gene knockout animals.
Moreover, in most mammalian species except rodents, no established ES cell lines are available that contribute efficiently to chimeric animals, which greatly limits the genetic studies in many species.
There are three potential alternative methods to generate gene-targeted animals in other species.
Approach I: Directly injecting DNA or mRNA of site-specific nucleases (ZNFs, TALENs, CRISPR/Cas system) into the one-cell embryo to generate DNA double-strand break (DSB) at a specified locus, which could drive both NHEJ-based gene disruption and homology directed repair (HDR)-based precise gene editing. My recent works have already showed we could used CRISPR/Cas system to generate mice carrying mutations in multiple genes and mice carrying reporter and conditional allele in one step. In future,I will focus on CRISPR/Cas system optimization (Specificity, Mosaicism and Efficiency) and genome engineering in large animals by CRISPR/Cas system.
Approach II: Establishing haploid ES cell lines from androgenetic haploid blastocysts, which partially maintain paternal imprints, and can be used as a genetically tractable fertilization agent for the production of live animals via injection into oocytes. My previous works shown that this procedure could be applied in mouse, and furthermore we could also generated haploid ES cells from Macaca fascicularis monkey parthenotes. In future, I will aim at producing androgenetic ES cells from primate and generate gene-modified primate by injecting these cells into oocytes.
Approach III: Cloning animals by injection of gene targeted nucleus of somatic cells or ES cells into enucleated oocyte to create gene-modified animals. Animals have been cloned in many species, but not in primate. However, NT-ES cells could be generated from cloned embryo from both primate and human. Therefore, I will concentrate on optimizing embryo culture and manipulation system and elucidating reprogramming mechanism to generate first cloned primate, which could be applied in genetically modified primate generation.
In summary, my interest is to apply different genome engineering approaches to generate genetically modified animals, especially primates, for modeling human disease and understanding gene function in development and disease.
姚璇 01 19179
周昌阳 02 19179
黄子健 02 19179
唐骋 01 19179
黄佳 02 19179
高妮 02 19179
贺冰冰 01 19179
李贺 01 19179
王兴 01 19179