Difference between revisions of "Team:BostonU"

Developing conditionally dimerizable split protein systems for genetic logic and genome editing applications

The field of synthetic biology seeks to engineer desirable cellular functionalities by developing molecular technologies that enable precise genetic manipulation. A promising solution is to reliably control proteins that naturally execute genetic modifications. Current strategies to regulate activity of such proteins primarily rely on modulating protein expression level through transcriptional control; however, these methods are susceptible to slow response and leaky expression. In contrast, strategies that exploit post-translational regulation of activity, such as conditional dimerization of split protein halves, have been demonstrated to bypass these limitations. Here, we compare the relative efficiency of previously characterized dimerization domains in regulating activities of three important genetic manipulation proteins - integrases and recombination directionality factors for genetic logic applications, and saCas9 for in vivo genome editing applications. We also establish guidelines to rationally identify promising protein split sites. Our characterization of these systems in mammalian cells ultimately paves way for important biomedical applications.