Researchers develop streamlined and efficient methods for genome editing
Researchers from the University of Copenhagen have further developed two of their previous gene-editing methods, have combined them and finally adapted them for CRISPR-Cas9 technology. The new methods package makes the editing process faster and more efficient for researchers investigating the function of genes. The study has recently been published in Nature Protocols.
In recent years genome editing has been developing rapidly. Especially genomic engineering using the CRISPR-Cas9 “DNA scissors” is currently the fastest growing research field in life sciences. The reasons for the “CRISPR craze” are obvious because the technology allows researchers to make any desired modification of the genome of any higher organism. By this virtue, CRISPR technology represents yet another small genomic revolution: while the past 2 decades of genome sequencing have revealed all the genes present in man and other organisms as well as a multitude of possible disease mutations, the CRISPR technology now enables individual researches to functionally study these genes and mutations by engineering them in cells or model animals in the laboratory.
FACS meets IDAA
The Nature Protocols paper describes two methods that make genome editing easier and that were developed by researchers from the Morten Frödin team at BRIC and the Eric Bennett team at Center for Glycomics, both at University of Copenhagen. The first method employs FACS enrichment of cells that express CRISPR-Cas9 which can dramatically increase or otherwise optimize editing efficiencies in a broad range of applications, as originally demonstrated by the Frödin team with Zinc Finger Nucleases (ZFNs) that preceded the more user-friendly and cost-efficient CRISPR system. The second method is called IDAA, which is a PCR-based analytical method that enables fast and sensitive detection of insertion or deletion (indel) mutations produced by CRISPR-Cas9 in cells, and it was originally developed by the Bennett team in collaboration with the Frödin group.
"In the Nature Protocols work we have further developed the FACS and IDAA methods for genome editing. Moreover, we have combined the two methods in a work flow, where they are used throughout a cell line engineering project, from the first step of screening new gRNAs to the final step of genotyping of your pools or clones of edited cells. Overall, this optimized pipeline can greatly reduce the workload of a cell line editing project," says Associate professor Morten Frödin.
The research of the Frödin group is supported by grants from the Danish Council for Independent Research/Medical Sciences, the Danish Cancer Society, the Novo Nordisk Foundation and the Lundbeck Foundation.
Genome editing using FACS enrichment of nuclease expressing cells and Indel Detection by Amplicon Analysis (IDAA), Nature Protocols (2017).