In a development that could accelerate the discovery of new diagnostics and treatments, researchers at Children's Hospital of Philadelphia (CHOP) have developed a versatile and low-cost technology for targeted sequencing of full-length RNA molecules. The technology, called TEQUILA-seq, is highly cost-effective compared to commercially available solutions for targeted RNA sequencing and can be adapted for different research and clinical purposes. The details are described in a paper in Nature Communications.
One way of doing so is using"long-read" RNA sequencing platforms, which sequence RNA molecules over 10,000 bases in length end-to-end, capturing the entirety of the transcript isoforms. However, these long-read platforms have modest sequencing yield, which has hampered their widespread adoption, especially in the clinical setting, as generating long-read RNA sequencing data at clinically informative depth could be prohibitively expensive.
One method that allows for targeted sequencing is called hybridization capture-based enrichment, which uses short pieces of nucleic acids called oligonucleotides as capture probes. These oligonucleotides are tagged with biotin molecules and designed to hybridize to their targets based on nucleic acid sequence complementarity, which allows for easy capture and isolation of their target sequences from a biological sample.
To benchmark its performance, the researchers performed TEQUILA-seq for multiple gene panels on synthetic RNAs or human RNAs. TEQUILA probes performed as well as commercial capture probes in target capture and enrichment, while being hundreds of times cheaper for each capture reaction. Moreover, the researchers demonstrated that TEQUILA-seq can substantially enhance detection while preserving quantification of target RNA molecules.
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