Efficient genome editing in rice with miniature Cas12f variants

Area of Science:

• Plant molecular biology
• Genome editing technologies
• Agricultural biotechnology

Background:

• CRISPR/Cas9 is widely used but has limitations like PAM requirements and delivery challenges.
• Hypercompact Cas12f (AsCas12f) from *Acidibacillus sulfuroxidans* is smaller than SpCas9 and targets T-rich motifs.
• AsCas12f exhibits low editing efficiency in plants, and its variants' performance is unknown.

Purpose of the Study:

• To evaluate the genome editing efficiency of AsCas12f variants in rice.
• To explore the potential of AsCas12f for targeted DNA deletion in plants.

Main Methods:

• Systematic investigation of genome cleavage activity of AsCas12f variants in rice.
• Analysis of editing efficiency and DNA modification patterns.

Main Results:

• Both AsCas12f-YHAM and AsCas12f-HKRA showed enhanced editing efficiency in rice.
• AsCas12f-HKRA achieved up to 53% editing efficiency.
• AsCas12f predominantly induced deletions, with a specific pattern and size range (10-11 bp).

Conclusions:

• AsCas12f variants, especially AsCas12f-HKRA, offer improved genome editing tools for plants.
• AsCas12f's unique deletion pattern shows potential for targeted DNA deletion applications.
• These findings expand options for precise genetic modification in agriculture.