UCSF researchers have developed a powerful Cas9 RNP-based technology that uses purified Cas9 ribonucleoproteins (RNP) for successful and efficient genome editing in primary human CD4+ T cells. Cas9 protein pre-complexed with a single guide RNA (sgRNA) is introduced as an RNP into human T cells by transient electroporation. The active complexes enabled the first successful Cas9-mediated homology directed repair (HDR) in primary human T cells. Cas9 RNPs have allowed generation of ‘knock-in’ primary human T cells with targeted genetic replacement of specific nucleotides, which was previously unattainable.
1) Unprecedented flexibility to ‘knock-out’ and ‘knock-in’ specific genetic elements in engineered T cells for cancer immunotherapy
2) New opportunity for therapeutic gene correction for primary immune deficiencies, treatment of infections and autoimmune diseases
3) Diverse research applications examining the function of coding and non-coding genetic variation in human immune regulation
Stage of Development
Proof of principle
In vitro human data