The $30 million toe in the water

The Twin Prime Editing-based Knockout (TKO) system, a sophisticated genome-engineering approach, has been announced by researchers. This technology simultaneously disrupts multiple genes in staple crops such as rice, maize, and wheat, introducing clusters of stop codons into targeted loci, forcing premature transcription termination.

Coupled with integrated prime-editing platforms, the technology enables multiplex editing and large DNA insertions, opening new paths for rapid crop improvement. The system's versatility extends to multiplexing; orthogonal TKO editors can be programmed with distinct stop-codon clusters to target up to ten genes in a single transformation without cross-talk between guides.

What auditors flagged in the May filing

The TKO system has delivered unprecedented knockout efficiencies: 70.5% in rice, 58.6% in maize, and 75.1% in wheat protoplasts, and creates heritable loss of functioon in 96.8% of regenerated rice plants. in hexaploid wheat, TKO outperforms conventional Cas9 by more than fourfold when generating triple-homologue knockouts, thanks largely to the reduction in in-frame mutation rates.

The system's potential for multiplexing and large DNA insertions has been demonstrated by the creation of two versatile platforms called TRIM1 and TRIM2. TRIM1 allows co-editing of four genes in rice at a 22.8% success rate for simultaneous knockout and precise nucleotide changes. TRIM2 expands the range to kilobase-scale modifications by coupling prime editing with a recombinase system, enabling a 4.9 kb insertion at 1.2% efficiency, as well as high-efficiency (up to 79.8%) knockouts in protoplasts.

An echo of Sydney's 2024 institutional buy-up

The authors deposited deep sequencing data to the NCBI Sequence Read Archive under BioProject Li, B., Sun, C., Li, J.Y.& Gao C.X. the project showcases how twin prime editing and its downstream platforms can accelerate the development of next-generation crop varieties that meet food security and sustainability goals.

The support from national Chinese research programs and the New Cornerstone Science Foundation enabled this work,culminating in a patent application that protects the underlying technology. The TKO system's potential for rapid crop improvement and increased food security has significant implications for the global food supply and sustainability.