Trial and error in CRISPR mediated genome editing: the devil is in the details.

BMC Seminar Thursday 11th of November, 12:00 in Læknagarður room 201

Speaker: Bylgja Hilmarsdóttir, Ph.D., clinical scientist at the molecular pathology unit, Department of pathology, Landspitali University hospital

Title: Trial and error in CRISPR mediated genome editing: the devil is in the details.

Zoom link: https://eu01web.zoom.us/j/63624676774

Abstract: The CRISPR/Cas technology has already revolutionized the field of genetic engineering and has multiple different applications in molecular biology. One of the most widely used CRISPR application is to make precise single nucleotide edits in the genome of cells. Although this is easy on paper the implementation of the method can be complex, depending on the experimental model used and the complexity of the desired gene editing.

In two CRISPR projects at our lab we have been using Cas9 and Cas12a to make precise genome editing in breast cancer cell lines. The aim of the talk is to discuss the practical aspects of using CRISPR for genome editing, using the two projects as examples. I will go over what has worked well in our hands, but maybe more importantly discuss major pitfalls and time-consuming optimization processes. Overall, the projects required a great deal of trial and error with some success in the end. I will discuss the efficiency of CRISPR/Cas genome editing in our lab both in regard to Cas9 and Cas12a. I will also talk about our experience in outsourcing the production of cancer cell line with precise gene edits. The hope is that our experience will be informative for others on the same path and inspire more collaboration among groups using CRISPR technology in Iceland.