Lecture with Professor Weizhi Ji
Mediated Gene Targeting in Monkeys with CRISPR/Cas9 and TALEN
Professor Weizhi Ji，
Yunnan Key Laboratory of Primate Biomedical Research，
Kunming Biomed International, Kunming, China
Place Hillarp lecture hall, Retzius väg 8, Solna Campus
Host Ken Chien, CMB and MedH
You are most welcome to attend the lecture but you must sign up for a seat in advance by emailing your name to Katarina.Drakenberg@ki.se
Professor Ji will present his recent work
TALEN-mediated gene mutagenesis in rhesus and cynomolgus monkeys.
Cell Stem Cell 2014 Mar;14(3):323-8
Generation of gene-modified cynomolgus monkey via Cas9/RNA-mediated gene targeting in one-cell embryos.
Cell 2014 Feb;156(4):836-43
Professor Ji got his PhD degree in Zoology at the Kunming Institute of Zoology in 1987 and completed a Post Doctoral Fellowship at the University of Oregon in 1989. In 1995 and 1997, he served as a visiting professor at the University of Wisconsin – Madison. After that, he served as the director of the Kunming Institute of Zoology. For the past 15 years, Prof. Ji’s research has focused on the reproductive biology of primates with the emphasis on in vitro culture of gametes. Part of the objective of this research is to develop and use modern techniques in reproductive technologies for making animal models for disease study and the conservation of endangered primates.
Nonhuman primates serve as important model species for studying human diseases and developing therapeutic strategies, yet the application of monkeys in biomedical researches has been significantly hindered by the difficulties in producing animals genetically modified at the desired target sites. Recent advances in gene editing technology, ZFNs, TALEN and CRISPR/Cas9 have introduced the potential for application of mutagenesis approaches in nonhuman primates to model human development and diseases. In the early of this year, we successfully applied the CRISPR/Cas9 and TALEN system, to target monkey genomes. In the CRISPR/Cas9 study, by coinjection of Cas9 mRNA and sgRNAs into one-cell-stage embryos, we successfully achieve precise gene targeting in cynomolgus monkeys. We also show that this system enables simultaneous disruption of three target genes (Nr0b1, Ppar-g and Rag1) in one step, and no offtarget mutagenesis was detected by comprehensive analysis. We also report successful TALEN-mediated mutagenesis of an X-linked, Rett syndrome (RTT) gene, methyl-CpG binding protein 2 (MECP2), in both rhesus and cynomolgus monkeys. Microinjection of MECP2-targeting TALEN plasmids into rhesus and cynomolgus zygotes leads to effective gene editing of MECP2 with no detected off-target mutagenesis. In summary, Both CRISPR/Cas9 and TALEN is an efficient and reliable approach for genetic modeling of human disease in nonhuman primates.Contact person: Katarina Drakenberg