CRISPR/Cas for PET Reporter Gene Engineering

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

DOI: 10.1007/978-1-0716-3499-8_17


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
SWORD Depositor: CSHL Elements
Depositing User: CSHL Elements
Date: 2024
Date Deposited: 20 Dec 2023 18:42
Last Modified: 09 Jan 2024 16:57
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