Cas12a knock-in Mice for multiplexed genome editing, disease modelling and immune-cell engineering

The pleiotropic effects of human disease and the complex nature of gene-interaction networks require knock-in mice allowing for multiplexed gene perturbations. We develop a series of knock-in mice of C57BL/6 background with conditional or constitutive expression of LbCas12a or high-fidelity enhanced AsCas12a, which were inserted at the Rosa26 locus. These Cas12a-based mouse lines, combined with the viral and non-viral delivery vehicles, provide a versatile toolkit for ex vivo and in vivo applications in genome editing, disease modelling, immune-cell engineering, and the deconvolution of complex gene interactions.