posted on 2021-03-18, 18:12authored bySuraj Makhija, David Brown, Rachel M. Rudlaff, Julia K. Doh, Struan Bourke, Yina Wang, Shuqin Zhou, Rasmi Cheloor-Kovilakam, Bo Huang
Recent
advances in genome engineering have expanded our capabilities
to study proteins in their natural states. In particular, the ease
and scalability of knocking-in small peptide tags has enabled high
throughput tagging and analysis of endogenous proteins. To improve
enrichment capacities and expand the functionality of knock-ins using
short tags, we developed the tag-assisted split enzyme complementation
(TASEC) approach, which uses two orthogonal small peptide tags and
their cognate binders to conditionally drive complementation of a
split enzyme upon labeled protein expression. Using this approach,
we have engineered and optimized the tag-assisted split HaloTag complementation
system (TA-splitHalo) and demonstrated its versatile applications
in improving the efficiency of knock-in cell enrichment, detection
of protein–protein interaction, and isolation of biallelic
gene edited cells through multiplexing.