Team:UIUC-Illinois

Why?

How?

Enzymatic Cascade

In utilizing the versatility of the PUF based toolkit we are developing, our current research involves an engineered metabolic pathway. The characterization and standardization of these genes in biobrick format is underway. Future goals involving the PUF toolkit will focus on tethering enzymes of productive artificial or natural pathways for kinetic favorability.

PUF

This is the PUF protein found on the PUM1 gene in humans. Our main focus and cause of interest in this protein is its ability to recognize single stranded RNA with specificity. The ability to modify PUF with tethered or fused proteins allows for the development of a customizable RNA binding toolkit. Depending on what fusion or tether is used, the functionality of the PUF protein is modified as well.

RNA scission

In fusion the PUF protein to a restriction endonuclease, the fusion protein demonstrates the ability to cleave single stranded RNA molecules with customizable specificity up to eight base pairs. Considering this ability, we believe PUF can prove itself as a promising new tool in researching and developing new types of gene therapy as it can silence genes with seqeunces specific to a reseachers interest.

RNA scaffold utilization

Since PUF has affinity towards RNA molecules, it is possible to synthesize or utilize existing stemloop structures in RNA in order to provide a kinetically favorable platform for tethered proteins to interact.