CRISPR/Cas9 Genetically-Modified Cells Exhibit NanoBRET-Donor Which Monitors Protein Interaction and Trafficking

Receptor-protein interaction and trafficking experiments are needed to study the operation of key drug targets, G-protein-coupled receptors (GPCRs). To understand such reactions and trafficking, a unique tool named bioluminescence resonance energy transfer (BRET) is used.

Previously, BRET has been restricted to the abnormal expression of labelled interaction partners. CRISPR/Cas9 genetic modification allows endogenous expression of luciferase-labelled proteins and thus overcomes the limitation.

CRISPR/Cas9-modified cells endogenously exhibiting a CXCR4/NanoLuciferase fusion protein in combination with β-Arrestin/Venus were utilized to observe receptor activation. Monitoring of receptor trafficking was permitted with the use of two fluorophores attached to a membrane and endosome standing CXCR4-reacting protein, respectively.

The Nluc BRET donor was successfully attached to endogenously-exhibited CXCR4 with the help of the novel CRISPR/Cas9 method. The resultant protein concentration was adequate to assess receptor interactions and internalization. The detection of the two acceptor fluorophores that are required for the internalization assay was made possible using the CLARIOstar monochromator.

Read the full article including the methods, results and discussion.

CRISPR/Cas9 genome-edited cells express nanoBRET-donor that monitors protein interaction and trafficking

About BMG Labtech

BMG LABTECH has been committed to producing microplate readers for more than twenty years. By focusing on the needs of the scientific community, the company’s innovative microplate readers have earned the company the reputation of being a technology leader in the field.

BMG LABTECH has developed a wide range of dedicated and multi-mode microplate readers for life sciences applications and high-throughput screening.

All BMG LABTECH microplate readers are "Made in Germany" and are conceived, developed, assembled, and tested entirely at our headquarters in Germany.

Since our establishment in Offenburg, Germany in 1989, BMG LABTECH has expanded to offer a worldwide sales and support network with offices in the USA, UK, Australia, Japan and France. Our subsidiaries, regional offices and distributors are committed to bringing you innovative microplate reader technology with the quality and reliability you expect from a German company.

Our staff includes engineers and scientists from the fields of biology, biochemistry, analytical chemistry, and physics.

Sponsored Content Policy: publishes articles and related content that may be derived from sources where we have existing commercial relationships, provided such content adds value to the core editorial ethos of News-Medical.Net which is to educate and inform site visitors interested in medical research, science, medical devices and treatments.

Last updated: Jul 30, 2018 at 10:34 AM

Other White Papers by this Supplier