Advanced Biocomputers in Mammalian Cells

Synthetic genetic circuits can revolutionize biotechnology, but engineering complex decision-making circuits remains challenging due to unintended interactions between cascading biological components.  We have previously developed a single-layered biocomputeration platform in mammalian cells called Boolean Logic and Arithmetic through DNA Excision (BLADE).  Here we will describe our recent efforts extending the BLADE platform to control guide RNA expression, thus allowing logic control of endogneous mammalian gene expression throught CRISPR/Cas.  We have also developed temporal logic circuits based on the BLADE platform.  These temporal logic circuits can generate at least five unique states from two inputs, and discriminate the order at which the two signals appear.  Together, these circuit designs will provide a new set of tools to integrate multiple features for enhancing tissue-specific trans- and endogenous gene regulation in animal models.