Chung, Taemoon, Merrill, Joseph R, Lyons, Scott K (2024) CRISPR/Cas for PET Reporter Gene Engineering. In: Methods in Molecular Biology. UNSPECIFIED, pp. 285-301. ISBN 9781071634981
Abstract
The relatively recent discovery of CRISPR/Cas has led to a revolution in our ability to efficiently manipulate the genome of eukaryotic cells. We describe here a protocol that employs CRISPR technology to precisely knock-in a PET imaging reporter transgene into a specific genetic locus of interest. Resulting transcription of the targeted reporter will more accurately mimic physiologic expression of the endogenous allele than conventional approaches, and so this method has the potential to become an efficient way to generate a new generation of "gold-standard" reporter transgenes. We break down the protocol into three experimental stages: how to identify the genomic location that the reporter transgene will be inserted, how to practically insert the reporter transgene into the genome, and how to screen resultant clones for the correct targeted event.
| Item Type: | Book Section |
|---|---|
| Subjects: | bioinformatics bioinformatics > genomics and proteomics > genetics & nucleic acid processing bioinformatics > genomics and proteomics Investigative techniques and equipment Investigative techniques and equipment > CRISPR-Cas9 bioinformatics > genomics and proteomics > genetics & nucleic acid processing > genomes |
| CSHL Authors: | |
| Communities: | CSHL labs > Lyons lab CSHL Cancer Center Program CSHL Cancer Center Program > Cellular Communication in Cancer Program |
| SWORD Depositor: | CSHL Elements |
| Depositing User: | CSHL Elements |
| Date: | 2024 |
| Date Deposited: | 20 Dec 2023 18:42 |
| Last Modified: | 29 Apr 2024 17:32 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/41331 |
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