Finding and Improving a Rat Virus Enzyme for More Efficient Prime Editing

Nineteen novel active reverse transcriptases were identified from a screening of 558 candidates, with enRERV-RT showing the highest activity.

• The optimized enRERV-RT variant outperforms conventional M-MLV-RT-based systems by 1.20-fold in mammalian and plant cells.
• At hard-to-edit loci, enRERV-RT provides a 1.88-fold increase in editing efficiency compared to M-MLV-RT.
• The high-throughput platform TRAP-seq-PE was developed to evaluate prime editor performance systematically.
• PE systems utilizing enRERV-RT demonstrate higher editing efficiencies across various mutation types than those using M-MLV-RT.
• The advancements may facilitate improvements in clinical gene therapy and crop breeding.

Simplified