{Reference Type}: Journal Article
{Title}: HUHgle: An Interactive Substrate Design Tool for Covalent Protein-ssDNA Labeling Using HUH-Tags.
{Author}: Smiley AT;Babilonia-Díaz NS;Hughes AJ;Lemmex ACD;Anderson MJM;Tompkins KJ;Gordon WR;
{Journal}: ACS Synth Biol
{Volume}: 13
{Issue}: 6
{Year}: 2024 Jun 21
{Factor}: 5.249
{DOI}: 10.1021/acssynbio.4c00188
{Abstract}: 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.