Aspects of the present disclosure are drawn to methods of improving the expression of secreted cuproenzymes from host cells by manipulating the expression level of one or more proteins involved in copper transport in the host cell, e.g., membrane-bound copper transporting ATPases and soluble copper transporters. The present disclosure also provides compositions containing such improved host cells as well as products derived from the improved host cells that contain one or more cuproenzymes of interest.

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.

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.

The present invention is related to a method to reduce peptide amidation activity in a given cell line, cell lines with reduced peptide amidation activity, and uses thereof.