A widely-applicable high-throughput cellular thermal shift assay (CETSA) using split Nano Luciferase

The use of cellular thermal shift assay (CETSA) to study Crizotinib resistance in ALK-expressing human cancers

Various forms of oncogenic ALK proteins have been identified in various types of human cancers. While Crizotinib, an ALK inhibitor, has been found to be therapeutically useful against a subset of ALK(+) tumours, clinical resistance to this drug has been well recognized and the mechanism of this phenomenon is incompletely understood. Using the cellular thermal shift assay (CETSA), we measured th...

Protein-protein interaction assay with split luciferase in planta

Comparative Analysis of Dual-Luciferase® Assay Technologies in a High Throughput Microplate Format

The importance of luciferase-based assays has been well documented over the past several decades1. With the advent of genetic engineering tools luciferase gene constructs are now routinely inserted into organisms or transfected into cells where they serve as reporters of gene activity2. This involves generation of a genetic construct where the luciferase gene is under the control of a transcrip...

Development of a high-throughput screening cancer cell-based luciferase refolding assay for identifying Hsp90 inhibitors.

The 90 kDa heat-shock protein (Hsp90) and other cochaperones allow for proper folding of nascent or misfolded polypeptides. Cancer cells exploit these chaperones by maintaining the stability of mutated and misfolded oncoproteins and allowing them to evade proteosomal degradation. Inhibiting Hsp90 is an attractive strategy for cancer therapy, as the concomitant degradation of multiple oncoprotei...

Identification of MMV malaria box inhibitors of plasmodium falciparum early-stage gametocytes using a luciferase-based high-throughput assay.

The design of new antimalarial combinations to treat Plasmodium falciparum infections requires drugs that, in addition to resolving disease symptoms caused by asexual blood stage parasites, can also interrupt transmission to the mosquito vector. Gametocytes, which are essential for transmission, develop as sexual blood stage parasites in the human host over 8 to 12 days and are the most accessi...