Genome Editing with the CRISPR/Cas System in Zebrafish

CRISPR/Cas is getting a lot of attention lately as a way to insert or delete genomic sequences in mammalian cells, but what about in model organisms? A new paper now shows that it works well in zebrafish and suggests it could be a new “go-to” method in other models as well.

Last month, we told you about the pros and cons of CRISPR/Cas compared to two more popular genome editing techniques, zinc finger nucleases (ZFNs) and transcription-activator-like effector nucleases (TALENs). To recap, the prokaryotic CRISPR/Cas system is kind of an immune system in bacteria and archaea that researchers have adapted to make changes in mammalian genomes. It works just as well or better than the other methods and is easier to do.

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Not much is known about CRISPR/Cas in model organisms, so the team from China tried it in zebrafish. Here’s what they found:

  • They could make site-specific indels in the zebrafish etsrp, gata4, and gata5 genes.
  • The frequency of indels was about the same as that reported for TALENs.
  • The phenotypes were what you’d expect for mutations in these genes, which suggested biallelic conversion.
  • They inserted mloxP precisely into the zebrafish genome, which they say shows great promise for making conditional knockouts in zebrafish, something that has been impossible until now.

“This work is one of the first two reports describing a highly efficient CRISPR system for genome editing in model organisms, which will facilitate cost-effective, genome-wide, functional analysis of coding and non-coding RNAs in vertebrate animals, including humans,” says Jing-Wei Xiong, Ph.D., who is at the Institute of Molecular Medicine at Peking University, and is one of the corresponding authors.

Find out if this could work for you at Cell Research, April 2013.