Nature biotechnologyJun 5, 2026

Precise multiple gene editing in monocot plants using paired prime editing tools

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jun 27, 2026

Abstract

Essence

Twin prime editing systems enabled multiplexed, precise knockout and editing in monocot crop models.

Evidence

This genome-engineering platform study tested TKO, TRIM1, and TRIM2 in rice, maize, and wheat protoplasts and regenerated rice plants, reporting knockout, coediting, and insertion efficiencies.

Caveat

Most efficiency results come from protoplast or plant-regeneration systems, so field performance and broad crop-trait effects are not assessed in the abstract.

Simplified

Simultaneously introducing diverse genomic edits remains a challenge in crop genome engineering. Here we describe a twin prime editing-based knockout (TKO) system that installs stop codon clusters (SCCs) for precise translational termination with minimal in-frame mutations. TKO achieves knockout efficiencies of up to 70.5%, 58.6% and 75.1% in rice, maize and wheat protoplasts, respectively, and produces heritable knockout alleles in 96.8% of regenerated rice plants. In hexaploid wheat, TKO outperforms Cas9 4.2-fold in generating triple-homolog knockouts, largely by reducing in-frame mutations. Orthogonal TKO editors with sequence-divergent SCCs enable simultaneous knockout of up to ten genes without cross-interference. Integration of TKO with conventional prime editing establishes TRIM1 (TKO editor-enabled gene rupture and development of integrated multitype genome modification system) for simultaneous knockout and precise editing, achieving a 22.8% coediting of four genes in rice. TRIM2 extends this capacity to kilobase-scale modifications through a prime editor-recombinase system, enabling a 4.9-kb insertion (1.2% efficiency) and gene knockout (up to 79.8%) in protoplasts.

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Precise multiple gene editing in monocot plants using paired prime editing tools

Abstract

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