Download Genome Editing in Human Stem Cells, Animals, and Plants

Genome Editing in Human Stem Cells, Animals, and Plants
Jin-Soo Kim
Center for Genome Engineering, Institute for Basic Science, Seoul, South Korea
Department of Chemistry, Seoul National University, Seoul, South Korea
Genome editing that allows targeted mutagenesis in higher eukaryotic cells and
organisms is broadly useful in biology, biotechnology, and medicine. We have
developed ZFNs, TALENs, and Cas9 RNA-guided endonucleases (RGENs), derived
from the type II CRISPR-Cas prokaryotic adaptive immune system, to modify
chromosomal DNA in a targeted manner. In particular, we used purified Cas9 protein
and in vitro transcribed guide RNAs rather than plasmids encoding these components
to correct large chromosomal inversions in the blood coagulation factor VIII gene that
cause hemophilia A in patient-derived induced pluripotent stem cells (iPSCs) and to
modify diverse genes in large animals and plants. The resulting animals and plants
contained small insertions or deletions (indels) at target sites, which are
indistinguishable from naturally-occurring variations, possibly bypassing regulatory
requirements associated with use of recombinant DNA.
Despite broad interest in RNA-guided genome editing, RGENs are limited by offtarget mutations. We developed Cas9 nuclease-digested whole genome (digenome)
sequencing (Digenome-seq) to profile genome-wide specificities of Cas9 nucleases in
an unbiased manner. Digenome-seq captured nuclease cleavage sites at single
nucleotide resolution and identified off-target sites at which indels were induced with
frequencies below 0.1%. We also showed that these off-target effects could be avoided
by using modified guide RNAs that contain two extra guanine nucleotides at the 5’ end.
Digenome-seq is a robust, sensitive, unbiased, and cost-effective method for profiling
genome-wide off-target effects of programmable nucleases including Cas9.
Education
1987 BS Dept. of Chemistry, Seoul National University
1989 MS Dept. of Chemistry, Seoul National University
1994 PhD Dept. of Biochemistry, University of Wisconsin-Madison
Experiences
Research Associate - Howard Hughes Medical Institute/MIT
1994 - 1997
Principal Investigator - Samsung Biomedical Research Institute 1997 - 1999
CEO and CSO - ToolGen, Inc. 1999 - 2005
Assistant/Associate Professor - Seoul National University 2005 - present
Director - Center for Genome Engineering, Institute for Basic Science 2014 - present
Jin-Soo Kim is an entrepreneur and chemist-turned-biologist. He graduated from Seoul
National University in 1987 with a major in chemistry. He then earned a master’s degree in
chemistry from Seoul National University in 1989 and a Ph.D. in biochemistry from the
University of Wisconsin-Madison in 1994. After postdoctoral training at Howard Hughes Medical
Institute/Massachusetts Institute of Technology, he came back to Seoul in 1997 to serve as
Principal Investigator at Samsung Biomedical Research Institute. He co-founded a
biotechnology company, ToolGen, Inc., focused on zinc finger technology in 1999, and served
as CEO and CSO for the subsequent 6 years. He joined the faculty of the Department of
Chemistry at Seoul National University in 2005. He now serves as Director of Center for
Genome Engineering at Institute for Basic Science. He has published over 60 articles and filed
20 patent applications, mostly in the field of gene regulation and genome editing. He has been a
member of Faculty of 1000 since May, 2013.
Throughout his independent scientific career, Jin-Soo Kim has been developing tools for
genome editing, a method now used widely in biomedical research, biotechnology, and
medicine. Genome editing enables genetic studies in model organisms and cell lines and gene
correction in stem and somatic cells for the treatment of both genetic and acquired diseases.
Furthermore, genome editing can be used to create value-added crops, livestock, fish, poultry,
and pets. The broad interest in this rapidly evolving and expanding technology among
researchers is highlighted by the choice of genome editing as the Method of the Year 2011 by
Nature Methods and as the Breakthrough of the Year runner-up by Science in 2013. Genome
editing in cultured cells, animals, and plants is catalyzed by programmable nucleases that
cleave chromosomal DNA in a targeted manner. The Kim group has developed and improved
three different types of programmable nucleases, namely, zinc finger nucleases (ZFNs), TAL
effector nucleases (TALENs), and RNA-guided engineered nucleases (RGENs) derived from
CRISPR/Cas, an adaptive immune system in prokaryotes. These tools are now used for
genome editing in human stem cells, model organisms, livestock, and plants in thousands of
labs all around the world.
The Kim group now focuses on target identification and validation for drug discovery
using genome-scale collections of TALENs and RGENs and on gene correction in human
pluripotent stem cells for the treatment of genetic disorders. He also collaborates with animal
scientists to develop genome-engineered pigs appropriate for organ transplantation and with
plant scientists to create value-added crops.