ExcScript Reverse Transcriptase

ExcScript Reverse Transcriptase (RT) was developed through in vitro evolution of M-MuLV RT. The enzyme possesses RNA-dependent and DNA-dependent polymerase activity but lacks RNase H activity, which is due to a mutation in the RNase H domain of M-MuLV RT. Compared to wild type M-MuLV RT, the engineered enzyme has dramatically improved thermostability, inhibition resistance, 50-fold higher processivity, robustness, and increased synthesis rate. The lack of RNase H activity allows ExcScript to produce long RNA transcripts of up to 20 kb. Additionally, the enzyme's high thermostability enables it to maintain full activity during the entire reverse transcription reaction and generate high yields of cDNA. The reaction temperature can be increased up to 65°C to efficiently transcribe RNA regions with high secondary structures or to improve specificity using gene-specific primers. The high processivity of ExcScript results in increased resistance to common reaction inhibitors. Highlights • Th