Free

Ribonucleoprotein delivery of Alt-R® CRISPR-Cas9 reagents for increased gen...

Event Information

Share this event

Date and Time

Location

Location

Life Sciences Institute, UBC

Room 161

Vancouver, BC V6T 1Z3

Canada

Event description

Description

High levels of genome editing using the CRISPR/Cas9 system can be achieved by optimization of key variables such as protospacer sequence, chemical modification of the guide RNA complex, and the delivery mode. Transfection of the CRISPR/Cas9 machinery as a ribonucleoprotein (RNP) complex has a number of benefits, such as high on-target potency combined with reduced off-target effects due to exposure control, and no observed toxicity or innate immune responses. Furthermore, a novel mutant Cas9 protein has been evolved to drastically reduce off-target gene editing while maintaining on-target potency. This mutant is ideally suited for RNP delivery allowing highly efficient genome editing without the complications associated with transfecting plasmid DNA. Strategies for optimization of CRISPR/Cas9 ribonucleoprotein complex delivery by improving electroporation conditions are discussed. Optimal electroporation parameters result in high levels of editing efficiency as well as high levels of cell viability.

About the speaker: Rolf Turk is a Staff Scientist at Integrated DNA Technologies in Coralville, Iowa. Rolf is originally from the Netherlands where he obtained a master’s degree in science at the University of Amsterdam. He continued his training as a graduate student at the Center for Human and Clinical Genetics at the Leiden University Medical Center. His research focused on the molecular mechanisms behind different forms of muscular dystrophy using gene expression profiling. After obtaining his Ph.D., he continued his research on muscular dystrophy as a post-doctoral researcher in the laboratory of Kevin P. Campbell, a HHMI Investigator at the University of Iowa. Since his employment at IDT in the fall of 2105, Rolf has been part of the Molecular Genetics Department where he focusses on the further development of RNA-guided endonucleases and optimizing delivery strategies.

Share with friends

Date and Time

Location

Life Sciences Institute, UBC

Room 161

Vancouver, BC V6T 1Z3

Canada

Save This Event

Event Saved