PUBLICATION
A Knock-in Zebrafish Reporter Line for Live Visualization of Endogenous Olig2 Protein Dynamics
- Authors
- Chang, C.T., Kawanishi, T., Nandagopal, S., Megason, S.G., Tsai, T.Y.
- ID
- ZDB-PUB-250925-4
- Date
- 2025
- Source
- Zebrafish : (Journal)
- Registered Authors
- Megason, Sean, Tsai, Tony
- Keywords
- CRISPR/Cas9, Olig2, knock-in, neural progenitor, pMN
- MeSH Terms
-
- Oligodendrocyte Transcription Factor 2*/genetics
- Oligodendrocyte Transcription Factor 2*/metabolism
- Genes, Reporter*
- Zebrafish*/genetics
- Zebrafish*/metabolism
- Animals
- Animals, Genetically Modified/genetics
- Animals, Genetically Modified/metabolism
- Gene Knock-In Techniques*
- Zebrafish Proteins*/genetics
- Zebrafish Proteins*/metabolism
- PubMed
- 40991403 Full text @ Zebrafish
Citation
Chang, C.T., Kawanishi, T., Nandagopal, S., Megason, S.G., Tsai, T.Y. (2025) A Knock-in Zebrafish Reporter Line for Live Visualization of Endogenous Olig2 Protein Dynamics. Zebrafish. :.
Abstract
The transcription factor oligodendrocyte transcription factor 2 (Olig2) plays a central role in specifying motor neurons and oligodendrocytes during vertebrate neural development. While transgenic reporter lines such as TgBAC(olig2:EGFP) have been instrumental in visualizing olig2 expression, they fall short in directly reporting endogenous protein levels and may not fully recapitulate native gene regulation. To address these limitations, we generated a TgKI(olig2-mNeonGreen) zebrafish line using CRISPR/Cas9-mediated knock-in at the endogenous olig2 locus. The resulting Olig2-mNeonGreen fusion protein localizes specifically to the nucleus, enabling direct live imaging and accurate quantification of Olig2-expressing cells. We confirmed that the knock-in preserves endogenous mRNA expression and protein function, and that homozygous fish develop normally. As proof of concept, modulation of Sonic Hedgehog signaling altered Olig2-mNeonGreen+ cell numbers as expected, confirming the reporter's responsiveness to known upstream inputs. This TgKI(olig2-mNeonGreen) line offers a robust tool for studying neural progenitor dynamics in vivo.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping