PUBLICATION
Protocol for generating a pericyte reporter zebrafish line Ki(pdgfrb-P2A-GAL4-VP16) using a CRISPR-Cas9-mediated knockin technique
- Authors
- Zi, H., Peng, X., Du, J., Li, J.
- ID
- ZDB-PUB-241215-1
- Date
- 2024
- Source
- STAR protocols 6: 103490103490 (Journal)
- Registered Authors
- Keywords
- CRISPR, Developmental biology, Model Organisms, Neuroscience
- MeSH Terms
-
- CRISPR-Cas Systems*/genetics
- Zebrafish*/genetics
- RNA, Guide, CRISPR-Cas Systems/genetics
- Receptor, Platelet-Derived Growth Factor beta/genetics
- Receptor, Platelet-Derived Growth Factor beta/metabolism
- Pericytes*/cytology
- Pericytes*/metabolism
- Blood-Brain Barrier/metabolism
- Animals
- Genes, Reporter/genetics
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- Animals, Genetically Modified
- Gene Knock-In Techniques*/methods
- PubMed
- 39673702 Full text @ STAR Protoc
Citation
Zi, H., Peng, X., Du, J., Li, J. (2024) Protocol for generating a pericyte reporter zebrafish line Ki(pdgfrb-P2A-GAL4-VP16) using a CRISPR-Cas9-mediated knockin technique. STAR protocols. 6:103490103490.
Abstract
Pericytes, the mural cells that envelop small blood vessels, play crucial roles in the formation of the blood-brain barrier (BBB). Here, we present a protocol for generating a pericyte reporter zebrafish line Ki(pdgfrb-P2A-GAL4-VP16) using a CRISPR-Cas9-mediated knockin technique. We describe steps for identifying efficient single guide RNA (sgRNA), constructing donor plasmid, and generating and maintaining the knockin line. We then detail procedures for in vivo imaging of brain pericytes. This protocol is adaptable for creating other knockin lines for specific cell labeling. For complete details on the use and execution of this protocol, please refer to Zi et al.1.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping