Abstract
We present a method of in vitro/in vivo protein detection by pairing CRISPR-Cas9 genome editing with the NanoBiT system. We describe steps for cell culturing, in vitro CRISPR-Cas9 ribonucleoprotein delivery, cell monitoring, efficiency assessments, and edit analysis through HiBiT assays. We then detail procedures to determine edit specificity through genomic DNA analysis, small interfering RNA reverse transfection, and HiBiT blotting. This protocol is simple to execute and multifunctional, and it enables high-throughput screens on endogenous proteins to be conducted with ease.
| Original language | English (US) |
|---|---|
| Article number | 103000 |
| Journal | STAR Protocols |
| Volume | 5 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jun 21 2024 |
Bibliographical note
Publisher Copyright:© 2024 The Authors
Keywords
- Biotechnology and bioengineering
- CRISPR
- Gene Expression
- Health Sciences
- High Throughput Screening
- Molecular Biology
- Protein Biochemistry
Fingerprint
Dive into the research topics of 'Protocol for HiBiT tagging endogenous proteins using CRISPR-Cas9 gene editing'. Together they form a unique fingerprint.Cite this
- APA
- Standard
- Harvard
- Vancouver
- Author
- BIBTEX
- RIS