Engineered dcas9 with reduced toxicity and its use in genetic circuits

Abstract

Disclosed herein are novel CRISPR/dCas9-based fusion proteins that facilitate the scaling up of genetic circuits, including those with non-linear response curves. The dCas9 based fusion proteins produce significantly less toxicity in comparison to previously described CRISPR/Cas9-based proteins used with logic gates. These improvements enable the generation of complex genetic circuits when both digital response curves and large amounts of dCas9 protein are needed. Also disclosed herein are methods of regulating expression of an output sequence through the introduction of novel CRISPR/dCas-9-based fusion proteins and genetic circuits into a cell.

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