Ultrashort‐Peptide‐Responsive Gene Switches for Regulation of Therapeutic Protein Expression in Mammalian Cells

Abstract Despite the array of mammalian transgene switches available for regulating therapeutic protein expression in response to small molecules or physical stimuli, issues remain, including cytotoxicity of chemical inducers and limited biocompatibility of physical cues. This study introduces gene switches driven by short peptides comprising eight or fewer amino acid residues. Utilizing a competence regulator (ComR) and sigma factor X‐inducing peptide (XIP) from Streptococcus vestibularis as the receptor and inducer, respectively, this study develops two strategies for a peptide‐activated transgene control system. The first strategy involves fusing ComR with a transactivation domain and utilizes ComR‐dependent synthetic promoters to drive expression of the gene‐of‐interest, activated by XIP, thereby confirming its membrane penetrability and intracellular functionality. The second strategy features an orthogonal synthetic receptor exposing ComR extracellularly (ComR EXTRA ), greatly increasing sensitivity with exceptional responsiveness to short peptides. In a proof‐of‐concept study, peptides are administered to type‐1 diabetic mice with microencapsulated engineered human cells expressing ComR EXTRA for control of insulin expression, restoring normoglycemia. It is envisioned that this system will encourage the development of short peptide drugs and promote the introduction of non‐toxic, orthogonal, and highly biocompatible personalized biopharmaceuticals for gene‐ and cell‐based therapies.