A stable enzyme is created by combining reverse transcriptase with a stabilizing protein, enhancing its durability and performance for DNA amplification methods like RT-PCR
Background
Reverse transcription is a cornerstone technique in molecular biology, facilitating the conversion of RNA into complementary DNA (cDNA) for various applications such as gene expression analysis, diagnostic testing, and the study of viral genomes. Methods like reverse transcription polymerase chain reaction (RT-PCR) depend on efficient and reliable reverse transcriptase enzymes to ensure accurate and sensitive detection of nucleic acids. As advancements in genetic research and clinical diagnostics continue to evolve, the necessity for robust and adaptable reverse transcription technologies becomes increasingly paramount to meet the growing demands for precision and high-throughput analysis.
Despite its critical role, current reverse transcriptase enzymes face significant challenges that impede their effectiveness in advanced applications. Many existing reverse transcriptases exhibit limited stability, particularly at elevated temperatures, which can reduce their enzymatic activity and shorten their usable lifespan. The purification processes for these enzymes are often complex and expensive, making them less accessible for widespread use.
Additionally, issues such as low solubility and sensitivity to harsh conditions further complicate their storage and handling, restricting their functionality under more rigorous experimental settings. These limitations hinder the scalability and efficiency of nucleic acid amplification methods, underscoring the need for improved enzyme solutions in the field.
Technology description
Stabilized reverse transcriptase fusion proteins comprise a thermostable reverse transcriptase enzyme linked to a stabilizer protein. This fusion enhances the overall stability of the enzyme, facilitating easier purification and increasing solubility. Additionally, the fusion allows for longer storage periods and enables the protein to function effectively under more demanding conditions, such as elevated temperatures. A flexible linker may be incorporated between the stabilizer protein and the reverse transcriptase to maintain proper structural configuration. These fusion proteins are ideally suited for applications in nucleic acid amplification techniques, including reverse transcription polymerase chain reaction (RT‑PCR) and various cDNA synthesis processes.
What sets these fusion proteins apart is their exceptional stability and resilience compared to traditional reverse transcriptases. The incorporation of a thermostable component ensures that the enzyme remains functional even at higher temperatures, which is crucial for efficient amplification and accurate results in molecular biology assays. The enhanced solubility and ease of purification reduce technical challenges and increase experimental reliability. Furthermore, the ability to store the proteins for extended periods without loss of activity makes them highly practical for laboratory use. These distinguishing features offer significant advantages, making the stabilized reverse transcriptase fusion proteins a superior choice for advanced genetic research and diagnostic applications.
Benefits
- Enhanced stability and longer storage life of reverse transcriptase enzymes
- Improved solubility and ease of purification of the fusion protein
- Ability to function effectively under higher temperatures and more rigorous conditions
- Facilitation of nucleic acid amplification methods such as reverse transcription PCR
- Versatility in cDNA synthesis applications
Commercial applications
- Reverse transcription PCR
- cDNA synthesis kits
- Molecular diagnostic assays
- Genetic research reagents
- Biotech amplification processes
Opportunity
The University of Texas at Austin is seeking an industry partner to license this technology.
Intellectual property
Issued patents: US 10,113,156; US.C1 10,150,955; US.D1 10,858,636; ZA 2011/07207; JP 5882064; EP 2403941; IL 214913; CH 2403941; DE 602010054054.5; FR 2403941; GB 2403941; NL 2403941.
Filed patents: CN 201080020123.1
