Fiche publication
Date publication
juillet 2025
Journal
Scientific reports
Auteurs
Membres identifiés du Cancéropôle Est :
Dr MARCHAND Virginie
Tous les auteurs :
Huber LB, Marchand V, Somtürk M, Müller S, Marx A
Lien Pubmed
Résumé
Reverse transcription polymerase chain reaction (RT-PCR) has evolved as a widely used approach in biotechnology and molecular diagnostics. It represents a powerful tool for amplifying and analysing RNA molecules and has therefore found widespread applications in profiling gene expression, viral detection and the diagnosis of various diseases. Wellestablished methodologies use viral reverse transcriptases (RTs) to transcribe RNA to cDNA and thermostable DNA polymerases (DNA pols) to amplify the resulting target sequence by PCR. This study reports on the development of novel Thermus aquaticus DNA polymerase I (Taq pol) variants that each are able to catalyse both steps simultaneously in a single tube without the need of viral RTs. In combination with their excellent thermostability (up to 95 °C), the novel Taq pol variants are suitable for employment in dye- or probe-based RNA detection methods. Moreover, the herein reported Taq pol variants are capable of performing multiplex detection of various RNA targets in a single tube with a single enzyme. Thus, discovery marks a significant advancement of current RT-PCR approaches and contributes simplifying and reducing costs in molecular diagnostics.
Mots clés
Taq Polymerase, genetics, Reverse Transcriptase Polymerase Chain Reaction, methods, Multiplex Polymerase Chain Reaction, methods, Protein Engineering, methods, Thermus, enzymology, DNA-Directed DNA Polymerase, genetics
Référence
Sci Rep. 2025 07 18;15(1):26147
