Optimized Virus-like Particles for Cas9 RNPs & Transgene/HDR Template Delivery

Tech ID: 32400 / UC Case 2021-177-0

Patent Status

Patent Pending

Brief Description

The inventors have developed optimized methods for using virus-like particles for the co-delivery of Cas9 ribonucleoprotein complexes and: 
  • a lentiviral genome that encodes a large transgene, such as a chimeric angtigen receptor (CAR) transgene 
  • a lentiviral genome that does not encode a sgRNA expression cassette 
  • a method for nucleofecting VLPs + homology directed repair (HDR) donor template together to enhance HDR in treated cells 


Suggested uses

In vivo/ex vivo generation of CRISPR-Cas9 gene edited CAR T cells.

Any application where a simultaneous gene knockout and transgene introduction/expression is desired.


This technology offers a one-step strategy using engineered lentiviral particles to introduce Cas9 RNPs and a CAR transgene into primary human T cells without electroporation. Furthermore, programming particle tropism allowed the inventors to target a specific cell type within a mixed cell population. This adaptable approach to immune cell engineering ex vivo provides a strategy applicable to genetic modification of targeted somatic cells in vivo.

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  • Doudna, Jennifer A.

Other Information

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