Evolved CRISPR-associated transposase systems for genome engineering

This technology is an evolved CRISPR-associated transposase system that facilitates highly efficient programmable DNA insertions in human cells without relying on homologous repair and double strand breaks.

Unmet Need: Large genetic insertion without inducing double-strand breaks

Current gene-editing technologies are either non-specific or inefficient and fail to integrate large genetic payloads without generating double-stranded DNA breaks or host homologous repair. Furthermore, while current methods can correct disease-causing mutations, regulatory approval and design of mutation-specific strategies significantly limit the number of patients who can benefit from therapeutic gene editing. There is a need for efficient targeted genomic integration of kilobase-scale payloads with minimal off-target events.

The Technology: RNA-guided DNA integration for programmable gene insertion

This technology is a CRISPR-transposon system that can facilitate kilobase-scale gene insertions with high specificity, minimal indels, and low off-target effects. Unlike current CRISPR/Cas9 systems, this technology uses Tn7-like transposons to catalyze RNA-guided DNA integration at defined sites without requiring double-strand breaks or host homologous repair. This technology has been optimized to enable multiplexed insertions, with integration events shown across multiple genomic targets in diverse hosts.

This technology has been extensively validated in bacterial cells and has shown efficacy in human cells, including primary human fibroblasts.

Applications:

• Gene editing therapy for genetically diverse patient populations
• Genetically engineered crops
• Generating models of human diseases (i.e., transgenic animal models)
• Cellular reprogramming
• Efficient site-specific DNA integration systems
• Large-scale multiplexed DNA insertion
• Bacterial genome engineering

Advantages:

• DNA integration without host homologous repair and double-strand break
• Integration of large genetic payloads
• Minimal indels and low off-target events

Lead Inventor:

Samuel Sternberg, Ph.D.

Patent Information:

Patents Issued (US 10,947,534, US 12,331,292)

Patents Pending (US 20220372521, US 20230147495, US 20240279629, US 20240209399, US 20250376701)

Related Publications:

• Witte IP, Lampe GD, Eitzinger S, Miller SM, Berríos KN, McElroy AN, King RT, Stringham OG, Gelsinger DR, Vo PLH, Chen AT, Tolar J, Osborn MJ, Sternberg SH, Liu DR. “Programmable gene insertion in human cells with a laboratory-evolved CRISPR-associated transposase.” Science. 2025 May 15;388(6748):eadt5199.

• Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH. “CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering.” Nat Biotechnol. 2021 Apr;39(4):480-489.

• Klompe SE, Vo PLH, Halpin-Healy TS, Sternberg SH. “Transposon-encoded CRISPR-Cas systems direct RNA-guided DNA integration.” Nature. 2019 Jul;571(7764):219-225.

Tech Ventures Reference:

• CU19096, CU21342, CU21338, CU21352, CU22030, CU23042, CU24383

• Licensing Contact: Cynthia Lang