Article
Custom CRISPR-Cas9 PAM variants via scalable engineering and machine learning.
Nature - 1 Jul 2025
Silverstein Rachel A, Kim Nahye, Kroell Ann-Sophie, Walton Russell T, Delano Justin, Butcher Rossano M, Pacesa Martin, Smith Blaire K, Christie Kathleen A, Ha Leillani L, Meis Ronald J, Clark Aaron B, Spinner Aviv D, Lazzarotto Cicera R, Li Yichao, Matsubara Azusa, Urbina Elizabeth O, Dahl Gary A, Correia Bruno E, Marks Debora S, Tsai Shengdar Q, Pinello Luca, De Ravin Suk See, Liu Qin, Kleinstiver Benjamin P
Abstract excerpt
Engineering and characterizing proteins can be time-consuming and cumbersome, motivating the development of generalist CRISPR-Cas enzymes1-4 to enable diverse genome-editing applications. However, such enzymes have caveats such as an increased risk of off-target editing3,5,6. Here, to enable scalable reprogramming of Cas9 enzymes, we combined high-throughput protein engineering with machine learning to derive...
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