Background
Reverse transcriptase enzymes are pivotal in molecular biology for transcribing RNA into DNA, facilitating studies from gene expression analysis to viral detection. The reliability and efficiency of reverse transcription are crucial for accurate cDNA formation, a foundational step necessary for subsequent applications such as cloning and quantitative PCR. Traditional reverse transcriptases, such as those derived from retroviruses, are often heat-labile and lack proofreading capability, leading to an increased likelihood of errors during transcription.
The limitations of current reverse transcription approaches include inadequate resistance to the heat needed to unwind RNA secondary structures, as well as a lack of exonuclease proofreading functions, which compromises the fidelity of the cDNA produced. Additionally, the existing tools exhibit a high error rate, further necessitating the development of more robust and accurate reverse transcriptase enzymes such as RTX.
Technology description
RTX is a robust reverse transcriptase enzyme engineered from KOD DNA polymerase. It features enhanced thermostability and an adept ability to precisely convert RNA into cDNA. RTX leverages structural modifications to better melt RNA structures, thereby facilitating the meticulous identification of RNA within biological samples. The enzyme retains the intrinsic proofreading exonuclease activity of its parent polymerase, which endows it with the ability to self-correct errant nucleotide incorporations.
The enzyme distinguishes itself from its counterparts by providing higher thermostability and potentially increased processivity, characteristics that underpin its more accurate representation of mRNA populations. This results in cDNA replication that is both high-fidelity and resilient to mutations. These traits, developed by the Lambowitz lab, make RTX particularly suitable for applications requiring precise RNA analysis.
Technologies
- Analyzing materials
- Compounds with saccharide radicals
- Enzymes
- Genetic testing
- Peptides
- Recombinant DNA
Benefits
- A high degree of thermostability enables RNA melting during reverse transcription.
- Proofreading function ensures high-fidelity cDNA synthesis.
- Improved enzyme processivity enhances comprehensive mRNA representation.
- Heat resistance allows for single-enzyme RT-PCR processes.
- Adaptability for various RNA analysis applications due to structural versatility
Commercial applications
- Quantitative PCR (qPCR) assays for gene expression studies
- High-throughput RNA sequencing and transcriptome analysis
- Diagnostic tests for RNA-based viruses including COVID-19 detection
- cDNA library construction for research and therapeutic development
- Molecular cloning applications necessitating accurate cDNA synthesis
Opportunity
The University of Texas at Austin is seeking an industry partner to license this technology.
