Chemical Control of Protease Activity and Gene Expression Using a Biotin-Released Inhibitory Domain

Chemical tools that enable precise temporal control of
protein
function are valuable reagents for probing dynamic processes within
live cells. Here, we introduce the biotin unblocking of the StrepTactin
steric block (BUSS) system, a novel chemogenetic tool that enables
precise temporal control of protein activity and interactions using
biotin. BUSS leverages the small StrepTagII (STII) peptide to flank
target domains, blocking their function by binding to StrepTactin,
with this steric block rapidly released upon biotin addition. Unlike
existing systems, which rely on larger protein tags that can disrupt
sensitive proteins, BUSS uses a smaller, minimally disruptive tag
compatible with a wide range of target proteins. We demonstrate the
versatility of the BUSS system for chemical control over diverse cellular
processes, including protein localization, protease activity, and
gene expression. With its compact design and broad utility, BUSS offers
a powerful approach for manipulating protein functions in living cells.