NSF Award
0109829
This Small Business Innovation Research (SBIR) Phase I project seeks to develop methods to prepare 60-200 base RNA/DNA oligonucleotides. Current methods for preparing long oligonucleotides suffer from limitations that hinder their application in biological science. Investigators have expressed an immediate need for longer material incorporating unnatural modifications and non-canonical substitutions. Access to these oligonucleotides is critical for continued discoveries in molecular biology and nucleic acid based therapeutics. Despite such demand, the most reliable synthetic method, 2-ACE chemistry, can provide quality oligonucleotides no longer than 50 bases. This SBIR Phase I project will use 2-ACE RNA to develop a novel biochemical technique using RNA ligase to enzymatically splice oligonucleotides together. RNA substrates will be coupled as part of a complex with a complementary splint. This project will evaluate different conformations of the ligation site, the tolerance of the enzyme for different substrate sequences, the optimal design and composition of the splint, and the ideal concentrations of substrates and cofactors. Once determined in an iterative process, the best conditions will be applied to more challenging research problems identified by collaborators. This research will directly address a critical deficit in the resources available to the national biotechnology research community.<br/><br/><br/>The commercial applications of this project are expected to be varied and immediate. As example, site-specifically modified RNAs of 50-200 bases in length should be in high demand by investigators who are developing RNA-targeted drugs or other RNA-dependent technologies beyond the reach of current synthetic means.
