The ability of the CRISPR-Cas9 gene editing system to enable high-specificity, site-specific double-stranded DNA breaks to effectively knockout a target gene, make precise base edits or stimulate gene activation or repression is continuing to revolutionize gene editing capabilities. With the use of fluorophore tags, Cas9-fusion proteins and Cas9 alternatives, this technology can be incorporated into a range of applications, from drug discovery screening approaches to monitoring protein interactions and multiplex engineering.
In our Gene Editing and CRISPR SelectScience Special Feature this month, we explore the latest technology and guidance in the field. You can learn from the experts by watching our webinars on demand, downloading application notes, viewing the videos, and reading key articles for the field of gene editing. In this article, we pull together some of the very hottest topics in the field today.
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Functional genomic screening exploits genomic editing to induce a measurable phenotypic change in a variety of organisms and tissues. This is broadly applicable to drug discovery workflows, from novel target discovery to target validation and better identification of compound mechanisms of action. While there are a number of screening platforms available, it’s CRISPR technology that is opening up new applications of genomic screening.
Bioluminescence resonance energy transfer (BRET) is a powerful tool for studying cellular protein interactions and trafficking. However, this technique is limited by the need for ectopic expression of labeled interaction partners.
In this application note, we look at the activation and internalization of membrane-spanning G protein-coupled receptors (GPCRs) using CRISPR/Cas9-edited fusion proteins. Using CRISPR/Cas9 technology, DNA coding for nanoluciferase (Nluc) is successfully inserted into the genomic locus of CXCR4 to act as an endogenously-expressed, BRET donor-labeled fusion protein. Coupled with suitable acceptor fluorophores, this can be used in receptor internalization assays, detected by the CLARIOstar from BMG LABTECH.
Learn how scientists at the UK's Institute for Genetic Medicine, in collaboration with the Wellcome Trust Centre for Mitochondrial Research, are using sophisticated consumable technologies to advance clinical research into rare mitochondrial diseases, paving the way towards major impacts on personalized medicine in mitochondrial and other rare diseases, as well as in cancer, Duchenne muscular dystrophy and an array of neurodegenerative disorders.
CRISPR-Cas9 technology can be incorporated into a range of screening approaches, from exploring drug mechanisms of action, to targeted drug resistance and identification of novel biomarkers. Depending on the expression effect required, there are three types of CRISPR screen available, which include:
In this application note by Horizon Discovery, we look at the combination of all three approaches (CRISPR KO, CRISPRi and CRISPRa) to identify components that influence resistance and sensitivity to the standard of care BRAF inhibitor, vemurafenib.
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