Nature communicationsFeb 18, 2026

Improved Un1Cas12f1 tool for editing multiple genes at once with better efficiency and range

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗OA ↗

Updated Jun 27, 2026

Abstract

Essence

An engineered Un1Cas12f1 variant expanded compact CRISPR targeting and improved editing activity.

Evidence

Bacterial library screening, mammalian cell validation, F0 mouse editing, transcriptional activation, and base-editing platform experiments showed evoCas12f recognized NTNR/NYTR PAMs, raised TTTR-site activity 1.4-fold, and reached up to 91% editing efficiency.

Caveat

The abstract reports platform performance across models rather than therapeutic delivery, off-target safety, or clinical outcome data.

Simplified

The compact CRISPR-Cas12f system is promising for AAV-delivered gene therapy, but its application has been constrained by restrictive PAM recognition (e.g., TTTR) and suboptimal editing efficiency. Through bacterial library screening and mammalian cell validation, we engineer evoCas12f, an optimized variant incorporating five key mutations, that dramatically expands PAM recognition to NTNR/NYTR. This advancement reduces median distance between two neighbouring PAM sites to 2 nucleotides in the human genome. It also demonstrates 1.4-fold enhanced activity at TTTR sites compared to wild-type Un1Cas12f1, achieving up to 91% editing efficiency. Remarkably, evoCas12f enables efficient generation of homozygous mutations in F0 generation mice, even at non-canonical PAM sites. We further adapt this system for robust transcriptional activation and precise base editing with a well-defined editing window. As a compact yet highly efficient platform, evoCas12f represents a significant advance in CRISPR technology, enabling multiplexed editing for high-resolution targeting applications and expanding possibilities for therapeutic genome engineering.