PUBLICATION

Generation of a zebrafish knock-in line expressing MYC-tagged Sox11a using CRISPR/Cas9 genome editing

Authors
Krueger, L.A., Morris, A.C.
ID
ZDB-PUB-220405-28
Date
2022
Source
Biochemical and Biophysical Research Communications   608: 8-13 (Journal)
Registered Authors
Morris, Ann C.
Keywords
CRISPR, Knock-in, Retina, Sox11, Zebrafish
MeSH Terms
  • Animals
  • CRISPR-Cas Systems
  • Gene Editing*/methods
  • Gene Knock-In Techniques
  • Recombinational DNA Repair
  • Zebrafish*/genetics
PubMed
35378361 Full text @ Biochem. Biophys. Res. Commun.
Abstract
Advances in CRISPR-Cas9 genome editing technology have strengthened the role of zebrafish as a model organism for genetics and developmental biology. These tools have led to a significant increase in the production of loss-of-function mutant zebrafish lines. However, the generation of precisely edited knock-in lines has remained a significant challenge in the field due to the decreased efficiency of homology directed repair (HDR). In this study, we overcame some of these challenges by combining available design tools and synthetic, commercially available CRISPR reagents to generate a knock-in line carrying an in-frame MYC epitope tag at the sox11a locus. Zebrafish Sox11a is a transcription factor with critical roles in organogenesis, neurogenesis, craniofacial, and skeletal development; however, only a few direct molecular targets of Sox11a have been identified. Here, we evaluate the knock-in efficiency of various HDR donor configurations and demonstrate the successful expression and localization of the resulting knock-in allele. Our results provide an efficient, streamlined approach to knock-in experiments in zebrafish, which will enable expansion of downstream experimental applications that have previously been difficult to perform. Moreover, the MYC-Sox11a line we have generated will allow further investigation into the function and direct targets of Sox11a.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping