Protocol for generating a pericyte reporter zebrafish line Ki(pdgfrb-P2A-GAL4-VP16) using a CRISPR-Cas9-mediated knockin technique
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.<sup>1</sup>.
Authors
- Xiaolan Peng (ORCID: https://orcid.org/0000-0001-9240-8510)
- Jiulin Du (ORCID: https://orcid.org/0000-0001-9413-9991)
- Huaxing Zi
- Jia Li (ORCID: https://orcid.org/0000-0002-2652-0034)
Institutions
- Chinese Academy of Sciences (CN)
- Center for Excellence in Brain Science and Intelligence Technology (CN)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-12-13
- DOI
- https://doi.org/10.1016/j.xpro.2024.103490
- Citations
- 3
- Primary Topic
- Zebrafish Biomedical Research Applications
- Type
- article
- Field-Weighted Citation Impact
- 0.98
Funders
- Natural Science Foundation of Shanghai
- National Natural Science Foundation of China
- Chinese Academy of Sciences
- Youth Innovation Promotion Association of the Chinese Academy of Sciences
- Youth Innovation Promotion Association