Zebrafish Endogenous Tagging via CRISPR Gene Editing
Tag your favorite gene at the native locus in your zebrafish line via CRISPR/Cas9 gene editing technology. Choose from a large selection of fluorescent proteins and epitope tags.
- Fluorescent tagging is typically used to aid in visualization of spatiotemporal dynamics of a given gene's protein product in live (and sometimes fixed) animals.
- Epitope tagging is most often utilized to facilitate biochemical studies, in particular for studies of protein-protein interactions which require the presence of a readily recognizable epitope or sequence feature for chemical modification or isolation.
(Left): SelN-BFPSTOP with BFP followed by a STOP codon inserted into the N-terminus of Selenon (SelN). (Center and Right): Ryr1a-mCherry line where the endogenous Ryr1a was tagged with mCherry at the N-terminus. Image courtesy of Melissa A. Wright, MD/PhD, Assistant Professor of Pediatric Neurology at University of Colorado.
Fluorescent Tagging vs. Epitope Tagging in Zebrafish
|Fluorescent Tagging||Epitope Tagging|
|Example of tags|
in fixed tissue samples
|Pros||Can aid in visualization of spatiotemporal dynamics of a given gene's protein product in live (and sometimes fixed) animals.||Can facilitate visualization or isolation of protein expression with
reduced chance of disrupting endogenous expression levels or
protein trafficking dynamics due to relatively small size.
Can also facilitate visualization or biochemical modification of proteins in fixed tissue with comparatively robust and cheap commercially available antibodies.
|Cons||Can interfere with gene function due to the relatively large size of fluorophores used.||Not sufficiently validated. Lack of publications.|
Zebrafish Endogenous Tagging Service Offerings
We can help you reduce experimental variability with our expression validation services through Western blot and/or immunostaining.
Custom tags and combinations (i.e. GFP::3xFLAG, etc.) are also available.