First-of-Its-Kind Study Characterizing the Mechanism of Template Switching With Ultraprocessive Reverse Transcriptase Enzymes

BRANFORD, Conn., August 27, 2024 (Newswire.com) - A new study demonstrates a significant advancement in our understanding of the template switching mechanism of reverse transcriptases (RTs) that is widely adopted to streamline RNA sequencing library preparation. The paper, Characterization and implementation of the MarathonRT template-switching reaction to expand the capabilities of RNA-Seq, just published in the RNA Society journal RNA, details the use of the UltraMarathonRT® (uMRT) enzyme and its progenitor, the MarathonRT®, to enhance the accuracy and efficiency of RNA sequencing, particularly for long or structured RNA transcripts. This breakthrough provides researchers with a powerful tool for capturing complete RNA sequences, offering deeper insights into RNA biology and expanding the potential for RNA research.

The study dissects the mechanisms behind template-switching reactions of RT enzymes used in RNA sequencing, a critical step for capturing full-length RNA sequences. Traditional methods have relied on distributive short-read RTs, such as MMLV RT enzymes. However, these methods have significant limitations due to the underpowered RT enzymes. The research, led by RNAConnect’s scientific co-founders, introduces a novel approach by adapting ultraprocessive RTs, derived from group II self-splicing introns, for use in RNA sequencing. The study provides a comprehensive understanding of the nucleotide specificity and the template-switching efficiency of the MarathonRT enzymes.

"This study represents a major milestone for RNA sequencing technology," said Li-Tao Guo, co-founder and VP of R&D at RNAConnect. "By leveraging the unique properties of our flagship product UltraMarathonRT, the team has described a more accurate and efficient method for RNA sequencing. This not only enhances our ability to analyze long and complex RNA molecules, but also opens new avenues for discovering RNA variants that were previously undetectable with conventional methods. Our goal is to continue pushing the boundaries of RNA research to unlock new insights into the complexities of RNA biology."

The new UltraMarathonRT-facilitated sequencing pipeline allows for the rapid and accurate sequencing of complex RNA libraries, including mixtures of long RNA transcripts. This method has been validated using both total human RNA and poly(A)-enriched RNA, showcasing its versatility and robustness in various RNA-seq applications. By optimizing each step of the RNA sequencing process from enzymatic reactions to template-switching oligonucleotide design, the study lays the foundation for a new era of RNA research, empowering scientists to explore the full diversity of RNA sequences and structures with unprecedented precision.

UltraMarathonRT Template Switching Kits designed to build RNA sequencing libraries based on this technology are available at www.rnaconnect.com.

About UltraMarathonRT Template Switching Kit:

The uMRT Template Switching Kit can provide full-length cDNAs in a simple reaction even when you do not know the sequences at the 5’ end of the RNA transcript. Its terminal transferase activity allows for template switching when used with the uMRT Template Switching Oligonucleotide. 

It is designed to capture low-abundance RNA with high efficiency and sensitivity. Kit components include uMRT, uMRT Buffer, uMRT Boost, uMRT Template Switching Oligonucleotide and the Template Switching Buffer.

About RNAConnect:

RNAConnect was founded by innovators in the RNA community, for innovators in the RNA community. By providing better enzymatic tools for visualizing and manipulating RNA, we can help our community discover, create, and innovate in ways previously not possible. For more information about RNAConnect and its offerings, to share your own ideas or to place your first order of uMRT, visit www.rnaconnect.com.

Source: RNAConnect