Fluorometric Assays for the Evaluation of Chemokine Receptor Inhibitors with Anti-HIV Activity
Poster Feb 14, 2007
Dominique Schols, Sigrid Hatse, Dana Huskens and Gary Bridger
In order to infect a target cell, the HIV envelope glycoprotein gp120 has to interact with both the cellular CD4 receptor and a chemokine receptor, CCR5 or CXCR4, the so-called HIV co-receptors. Several flow cytometric/fluorometric assays are developed in our lab and are very helpful in deciphering the mode of action and interaction site of several classes of HIV entry inhibitors, such as the chemokine receptor antagonists.
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