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[Seminar Notification] Dr. Jin-Soo Kim_12.08.2016
2016-11-30

1. Title : Genome Editing in Human Stem Cells, Animals, and Plants

2. Speaker : Jin-Soo Kim, Ph.D.

3. Affiliation : Center for Genome Engineering, Institute for Basic Science, Seoul, South Korea Department of Chemistry, Seoul National University, Seoul, South Korea

4. Date : 12. 08. 2016 16:00-17:00

5. Place : SIMS Main Building, Auditorium 109 (1F)

6. Abstract :

Genome editing that allows targeted mutagenesis in cells and organisms is broadly useful in biology, biotechnology, and medicine. We have developed ZFNs, TALENs, and Cas9/Cpf1 nucleases to modify chromosomal DNA in a targeted manner. In particular, we used purified Cas9/Cpf1 proteins rather than plasmids to correct large chromosomal inversions in the factor VIII gene that cause hemophilia A in patient-derived iPSCs or to modify diverse genes in animals and plants. The resulting cells and organisms contained small 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, Cas9 and Cpf1 are limited by off-target mutations. We developed nuclease-digested whole genome sequencing (Digenome-seq) to profile genome-wide specificities of Cas9 and Cpf1 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 purified Cas9/Cpf1 ribonucleoproteins (RNPs) and modified guide RNAs. Digenome-seq is a robust, sensitive, unbiased, and cost-effective (< USD 1,500) method for profiling genome-wide off-target effects of programmable nucleases.