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Technology Seminar (10 Jun 2016)

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Technology Seminar

Rapid and Highly Efficient Mammalian Cell

 

Engineering via Cas9 Protein Transfection 

 By

Dr Xiquan Liang

Staff Scientist from Molecular Biology Business Unit, USA

 

10 Jun 2016 (Fri)

3 - 4 pm

 

Seminar Room 1A, G/F

The Hong Kong Jockey Club Building for Interdisciplinary Research

5 Sassoon Road, Pokfulam, Hong Kong

 

Abstract:

CRISPR-Cas9 systems provide an innovative solution for high efficiency genome editing.  Via Cas9 protein transfection, we achieved indel rates of up to 94% in Jurkat T cells and 87% in induced pluripotent stem cells (iPSC) for a single target, and approximately 93% and 65% for two and three targets, respectively.  Recently, we improved the design and delivery of gRNA, Cas9 nuclease, and donor DNA.  Under optimal conditions, we achieved approximately 52% targeted integration efficiency for one nucleotide substitution and 40% efficiency for six nucleotide insertion in HEK293 reporter cells.   In addition, we report precise genome editing rates of up to 24% in induced pluripotent stem cells (iPSCs) and 40% in Cas9-expressing iPSCs for a single nucleotide substitution at multiple genomic loci.  Taken together, we provide simple and highly efficient approaches for modulation of the mammalian genome and for generation of knock-in and knock-out cell lines.


Speaker:

 Dr Xiquan Liang graduated from City College of New York in 1999. He joined Thermo Fisher Scientific as a staff scientist (2004).  Since then he has led development of product development leadership in protein analysis, including High Molecular Weight MS standard, SILAC for protein identification and quantification, and Dynabeads-TiO2 for phosphopeptide enrichment.   He is now engaged in product development in Synthetic Biology, including Bluegrass, GeneArt seamless DNA assembly tools, site-directed mutagenesis, and CRISPR-mediated mammalian cell engineering in the Life Sciences Solutions Group of Thermo Fisher Scientific in Carlsbad, CA.  He is focused on developing tools for the entire Synthetic Biology workflow, specifically Bioinformatics, Gene Synthesis and Assembly, and Molecular and Cell Engineering.”

 

ALL ARE WELCOME


Co-organized with Life Technologies. Euro Tech (Far East) Ltd Logo
Refreshment to be served

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