Production and functional testing of probes mimicking human sperm CRISP1, CRISP2, and TMEM95 for oocyte receptor identification

The sperm proteins CRISP1, CRISP2, and TMEM95 are crucial for human gamete fusion, yet their oocyte interactors remain unidentified. To achieve this goal, we are producing high-avidity pentamerized probes mimicking these three proteins in Expi293F cells in order to assess their ability to bind to oocytes. Moreover, for CRISP2, an in-silico screening of the human oocyte surfaceome previously performed in our lab identified candidate oocyte interactors. To validate these results, HeLa cells expressing these proteins are incubated with the probes and CRISP-derived synthetic peptides and the binding between the probes and the cells is evaluated in immunofluorescence.