Following our work on synthetic long noncoding RNA (slncRNA) scaffolds, we have begun to incorporate RNA-binding proteins into the scaffolds. We utilize slncRNAs containing phage coat-protein binding sites, and their cognate RNA-binding coat proteins, to form nucleoprotein granules, both in vitro and in vivo. Using fluorescent labeling on the slncRNA and proteins, we can track characterize the granules by tracking the fluorescence signal over time. We are currently exploring different applications based on these granules:
- Granules constitute a genetically encoded storage compartment for protein and RNA, with a programmable controlled release profile that is determined by the number of hairpins encoded into the RNA. This has uses for medical applications.
- We can encode multiple types of binding sites into the scaffold, which bind multiple RNA-binding enzymes. This may improve the efficiency of enzymatic reactions.
- Scaffolds containing synthetic RNA-binding human cell receptors can act as broad-spectrum decoy particles for viruses.
Relevant publications:
Phase separation-based antiviral decoy particles as basis for programmable broad-spectrum therapeutics
Or Willinger, Naor Granik, Sarah Goldberg and Roee Amit. BioRxiv.
GelMetrics: An algorithm for analyzing the dynamics of gel-like phase separated condensates
Naor Granik and Roee Amit. BioRxiv.
A Cell-Free Assay for Rapid Screening of Inhibitors of hACE2-Receptor–SARS-CoV-2-Spike
Nanami Kikuchi, Or Willinger, Naor Granik, Reut Gal, Noa Navon, Shanny Ackerman, Ella Samuel, Tomer Antman, Noa Katz, Sarah Goldberg and Roee Amit. ACS Synthetic Biology 11, 4, 1389–1396 (2022).
Formation of synthetic RNA protein granules using engineered phage-coat-protein -RNA complexes
Naor Granik, Noa Katz, Or Willinger, Sarah Goldberg and Roee Amit. Nature Communications 13, 6811 (2022).