CRISPR-transposon platform for programmable DNA integration

This technology is an RNA-guided genome engineering platform that enables precise, site-specific DNA integration and endogenous protein tagging without introducing double-strand breaks.

Unmet Need: Programmable genomic tools for DNA modification and gene targeting

Current genome engineering approaches primarily rely on nuclease-based systems that create double-strand DNA breaks, which are inefficient and highly cell-state-dependent. These platforms can introduce unintended mutations and genomic instability, which can often result from imprecise insertion of large genetic payloads or multiplexed edits. As a result, there is a need for more reliable strategies that enable accurate, site-specific DNA integration while minimizing cellular damage. Addressing these limitations is essential for advancing translational applications that require durable and predictable genome modifications.

The Technology: CRISPR-based transposon platform for genome integration

This technology enables targeted DNA integration by coupling RNA-guided nucleic acid modification with transposase-mediated insertion, allowing genetic material to be placed at defined genomic locations. A guide RNA directs a CRISPR-associated protein complex to a specific target sequence in the genome, where transposon-associated proteins then catalyze the insertion of a genetic payload. The payload is flanked by engineered transposon end sequences that are optimized to promote precise integration. By avoiding reliance on homology-directed repair, the approach supports insertion of large DNA cargos and enables applications in gene addition and endogenous protein tagging.

This technology has been validated with pooled donor DNA libraries.

Applications:

• Targeted DNA integration and gene addition
• Endogenous protein tagging
• Large-cargo and multiplexed genome engineering
• Research and discovery platforms for studying genome function

Advantages:

• Enables precise, site-specific DNA integration
• Reduces unintended mutations associated with nuclease-based genome editing
• Supports insertion of large DNA cargos
• Improves predictability and durability of genome insertions
• Allows programmable targeting using guide RNAs
• Enables endogenous protein tagging and multiplexed genome modifications

Lead Inventor:

Samuel H. Sternberg, Ph.D.

Patent Information:

Patent Pending

Related Publications:

• Gelsinger DR, Vo PLH, Klompe SE, Ronda C, Wang H, Sternberg SH. “Bacterial genome engineering using CRISPR RNA-guided transposases.” bioRxiv. 2023 Mar 21.

• Walker MWG, Klompe SE, Zhang DJ, Sternberg SH. “Novel molecular requirements for CRISPR RNA-guided transposition.” Nucleic Acids Res. 2023 May 22; 51(9): 4519-4535.

Tech Ventures Reference:

• IR CU22373, CU23110, CU23204

• Licensing Contact: Cynthia Lang