Stem Cell Genetics

Kosuke Yusa, Ph.D. Professor btn

A variety of genes are involved in every biological process. Identifying these genes and understanding their biological functions are the basis of biology and clinical translation of research outcome. Applying genome editing technologies, our laboratory developed a powerful forward genetic screening method – CRISPR screening – and has provided toolkits to the research community. Our current research aims to understand the molecular mechanisms underlying proliferation and differentiation of cancer and pluripotent stem cells using CRISPR screening, and develop novel therapeutic applications.

Research and Education

To understand gene functions using genetics, we identify causal relationship between phenotype and genotype. While reverse genetics focuses on genes of interest and analyze phenotypes of those mutants, forward genetics provides a powerful mean to comprehensively identify genes involved in a phenotype of interest. However, methods available for studies using mammalian cells were not as high as hoped. To overcome this limitation, we applied CRISPR-Cas9 systems and developed powerful ‘CRISPR screening’.

Our current research interest is to understand the molecular mechanism by which cancer cells and pluripotent stem cells proliferate and differentiate, and we take advantage of our CRISPR screen technique to start addressing new biological questions. We have performed a large-scale CRISPR screens on a panel of 234 cancer cell lines and catalogued fitness genes, from which novel molecular target therapy could be developed. For pluripotent stem cells, we are particularly interested in the variation seen amongst ES/iPS cell lines. Understanding the mechanisms would allow us to develop robust differentiation protocols and facilitate the translation of pluripotent cell technologies.

Figure 1. Workflow of CRISPR screening
A mutant cell library can be subjected to various type of phenotypic analysis.

 

Figure 2. Colorectal cancer cell lines
Each cell line shows distinct morphology. By screening these cell lines, we identified that the WRN gene is specifically required for cells with microsatellite instability.


Recent Publications

  1. Behan FM, Iorio F, Picco G, Goncalves E, Beaver CM, Migliardi G, Santos R, Rao Y, Sassi F, Pinnelli M, Ansari R, Harper S, Jackson DA, McRae R, Pooley R, Wilkinson P, Meer D, Dow D, Buser-Doepner C, Bertotti A, Trusolino L, Stronach EA, Saez-Rodriguez J, Yusa K, Garnett MJ. Prioritisation of oncology therapeutic targets using CRISPR-Cas9 screening. Nature 568:511-516 (2019)
  2. Koike-Yusa H, Li Y, Tan EP, Velasco-Herrera MD, Yusa K. Genome-wide recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library. Nature Biotechnol. 32:267-273 (2014)
  3. Yusa K, Rashid ST, Strick-Marchand H, Varela I, Liu PQ, Paschon DE, Miranda E, Ordóñez A, Hannan NR, Rouhani FJ, Darche S, Alexander G, Marciniak SJ, Fusaki N, Hasegawa M, Holmes MC, Di Santo JP, Lomas DA, Bradley A, Vallier L. Targeted gene correction of a1-antitrypsin deficiency in induced pluripotent stem cells. Nature 478: 391-394 (2011)
  4. Yusa K, Zhou L, Li MA, Bradley A, Craig NL. 2011. A hyperactive piggyBac transposase for mammalian applications. Proc. Natl. Acid. Sci. USA 108: 1531-1536 (2011)
  5. Yusa K, Horie K, Kondoh G, Kouno M, Maeda Y, Kinoshita T, Takeda J. Genome-wide phenotype analysis in ES cells by regulated disruption of the Bloom’s syndrome gene. Nature 429: 896-899 (2004)

Stem Cell Genetics

Professor: Dr Kosuke Yusa
Assistant Professor: Dr Yusuke Tarumoto
Assistant Professor: Dr Gohei Nishibuchi
Assistant Professor: Dr Kazunari Aoki
Assistant Professor: Dr Fumihiko Kawamura
TEL: +81 75 751 4100
e-mail: k.yusa@infront.kyoto-u.ac.jp