Connecting multiple tRNA molecules, including in vitro transcribed as well as native tRNAs, to enable sequencing and structural applications
Background:
Transfer RNA (tRNA) molecules play a crucial role in biological processes, but technical limitations hinder their analysis. The short length of individual tRNAs makes them challenging to sequence effectively using current high-throughput sequencing technologies, which are optimized for longer nucleic acid molecules. Additionally, the complex secondary and tertiary structures of tRNAs, along with extensive post-transcriptional modifications, create further obstacles for comprehensive analysis. These limitations restrict our understanding of tRNA biology, modifications, and their roles in various cellular processes and diseases.
Technical Overview:
Northeastern researchers have developed an innovative strategy for connecting multiple tRNA molecules into longer chains using enzymatic or chemical linkage methods. This approach overcomes the limitations of short tRNA length by creating extended constructs that are more amenable to high-throughput sequencing platforms. The method preserves tRNA integrity and modifications while enabling more efficient sequencing and analysis. The linking strategy can be applied to both native tRNAs isolated from cells and in vitro transcribed tRNAs, providing flexibility for different research applications.
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