High-throughput method for gene insertion

This technology is a high-throughput method to introduce single or multiple genetic insertions in human cell lines.

Unmet Need: Large-scale implementation of genetic insertions

Despite recent breakthroughs in CRISPR/Cas9 genome editing technologies, the implementation of CRISPR/Cas9 genome editing technologies in human and additional mammalian cell lines has remained cumbersome and challenging. Thus, tools that manipulate the genome of human cell lines in a massively parallel and efficient manner are needed for multiple areas of biomedical research.

The Technology: High-throughput method to create parallel genetic alterations

This technology is a vector-based system that has the capacity to modify a plurality of target sites in the mammalian cell. By using multiple Cas9 proteins to prevent competition, this technology creates libraries with single transgene populations for mammalian cell lines. The system is designed for incorporation into parallelized, high-throughput genome editing workflows, which were previously limited to bacteria and yeast.

This technology has been validated with human cell lines.

Applications:

• High-throughput generation of transgenic animal models
• Somatic cell gene therapy
• Precision drug screening
• Somatic cell therapeutic implantation

Advantages:

• Enhances Cas9 activity
• High-throughput generation of isogenic human cell lines
• Generates genetic perturbations in parallel
• Rapidly integrates defined genetic sequences into a target genome with high specificity

Lead Inventor:

Alejandro Chavez, M.D., Ph.D.

Patent Information:

Patent Pending (US20220275400)

Related Publications:

• Lampe GD, King RT, Halpin-Healy TS, Klompe SE, Hogan MI, Vo PL, Tang S, Chavez A, Sternberg SH. “Targeted DNA integration in human cells without double-strand breaks using CRISPR-associated transposases.” Nature biotechnology. 2024 Jan;42(1):87-98.

Tech Ventures Reference:

• IR CU19411

• Licensing Contact: Cynthia Lang