Cas12a/Cpf1 knock-in mice enable efficient multiplexed immune cell engineering
Dong, M. B.; Tang, K.; Zhou, X.; Shen, J.; Chen, K.; Kim, H. R.; Zhou, J.; Cao, H.; Vandenbulcke, E.; Zhang, Y.; Chow, R. D.; Du, A.; Suzuki, K.; Fang, S.-Y.; Majety, M.; Dai, X.; Chen, S.
Show abstract
Cas9 transgenic animals have drastically accelerated the discovery of novel immune modulators. But due to its inability to process its own CRISPR RNAs (crRNAs), simultaneous multiplexed gene perturbations using Cas9 remains limited, especially by pseudoviral vectors. Cas12a/Cpf1, however, can process concatenated crRNA arrays for this purpose. Here, we created conditional and constitutive LbCas12a knock-in transgenic mice. With these mice, we demonstrated efficient multiplexed gene editing and surface protein knockdown within individual primary immune cells. We showed genome editing across multiple types of primary immune cells including CD4 and CD8 T cells, B cells, and bone-marrow derived dendritic cells. These transgenic animals, along with the accompanying viral vectors, together provide a versatile toolkit for a broad range of ex vivo and in vivo gene editing applications, including fundamental immunological discovery and immune gene engineering.
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