@article {70380, title = {Improved sensors for fructose-1,6-bisphosphate enable in vivo imaging of glycolysis}, journal = {bioRxiv}, year = {2026}, month = {2026 May 01}, abstract = {

Fructose-1,6-bisphosphate (FBP) is the product of the first committed step of glycolysis, and its concentration is tightly correlated with glycolytic flux. Glycolytic activity varies across tissues and cell types: some tissues, such as the brain, dynamically regulate glycolysis in response to demand, while others, such as the liver have characterized spatial heterogeneity. Here, we report HYlight2, an improved sensor for FBP developed through random whole-gene mutagenesis in E. coli lysate. After four rounds of screening, we isolated HYlight2, which retains its binding affinity while displaying a ΔR/R \~9 in vitro, a three-fold improvement in mammalian cells, and a two-fold improvement in detecting glycolytic responses during stimulated neuronal activity. We further demonstrate its use in vivo to detect altered glycolytic activity in C. elegans neurons, zebrafish pancreatic islets, and mouse liver.

}, doi = {10.64898/2026.04.29.721630}, url = {https://www.biorxiv.org/content/early/2026/05/01/2026.04.29.721630}, author = {Tyler, Juliann and Vishwanath, Anjali Amrapali and Menon, Triveni and Duarah, Tanishqua and Adhikari, Raghabendra and Koberstein, John N. and Feliciano, Daniel and Espinosa-Medina, Isabel and Col{\'o}n-Ramos, Daniel and Tebo, Alison G.} }