HUHgle: An Interactive Substrate Design Tool for Covalent Protein-ssDNA Labeling Using HUH-Tags

HUH-tags have emerged as versatile fusion partners that
mediate
sequence specific protein-ssDNA bioconjugation through a simple and
efficient reaction. Here we present HUHgle, a python-based interactive
tool for the visualization, design, and optimization of substrates
for HUH-tag mediated covalent labeling of proteins of interest with
ssDNA substrates of interest. HUHgle streamlines design processes
by integrating an intuitive plotting interface with a search function
capable of predicting and displaying protein-ssDNA bioconjugate formation
efficiency and specificity in proposed HUH-tag/ssDNA sequence combinations.
Validation demonstrates that HUHgle accurately predicts product formation
of HUH-tag mediated bioconjugation for single- and orthogonal-labeling
reactions. In order to maximize the accessibility and utility of HUHgle,
we have implemented it as a user-friendly Google Colab notebook which
facilitates broad use of this tool, regardless of coding expertise.