CRISPR mutants and morphants generation

The zebrafish is a powerful tool for in vivo functional characterization of gene function and validation of candidate genomic sequences.

 

Until recently most loss-of-function studies in zebrafish have heavily relied on knocking-down target gene expression via morpholinos. By binding to complementary RNA, morpholinos antisense oligomers have been shown to be effective in knocking down some genes up to 5 days post-fertilization when injected into one cell stage embryos. Although MO knocking down continue to be a valuable tool for the field it has also obvious limitations such as a lack of heritability of the morphants as well as by causing some non-specific off target effects.

 

To have an unambiguous inside of the phenotype, some researchers prefer technologies allowing the generation of mutants. Given the ease of use of current site-specific nuclease technologies, the CRISPR system has demonstrated unprecedented efficacy and facility in enabling targeted gene disruption in zebrafish.

 

We offer both services: morpholinos provide, in principle, and accessible and straightforward method for gene knockdown in zebrafish.

But as some publications (see here and here) show discrepancies between morphants and mutants, we strongly advise you to confirm the phenotype by generating mutants with the CRISPR technology.

 

For more information, to discuss your needs, or for a quote, please contact us.