Transcreener™ PDE Assay: Homogenous AMP and GMP Detection
Poster Mar 13, 2007
Matt Staeben, Rebecca Schall Josvai, Thane A. Westermeyer, Tony A. Klink, Jennifer A. Fronczak, and Karen M. Kleman-Leyer
Phosphodiesterases (PDEs) control a myriad of cellular processes through their ability to hydrolyze the second messenger signaling molecules, cAMP and cGMP. This superfamily, which is grouped into 11 different gene families generating >50 different forms of PDE, has been recognized as drug targets in the therapeutic treatment of congestive heart failure, thrombosis, hypertension, depression, and erectile dysfunction.
To facilitate the rapid identification of PDE inhibitors, BellBrook Labs has developed a fluorescence polarization immunoassay that directly detects AMP and GMP. The Transcreener™ PDE Assay includes a single set of reagents - an anti-AMP/GMP antibody and fluorescent tracer - that can be used for any cyclic nucleotide PDE, providing a robust, cost-effective alternative to currently available technologies. The use of a far red tracer minimizes the potential for compound interference. Here, we describe the development of the Transcreener™ PDE Assay and its application in identifying compounds that inhibit their activity
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE