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Abstract Achieving coordinated CRISPR-based transactivation (CRISPRa) regulation in vivo remains challenging, as the requisite genetic elements frequently exceed the packaging capacity of AAV vectors. Here we report the identification and engineering of the Cas12m protein family, a naturally nuclease-dead CRISPR system that possesses intrinsic self-processing activity for its precursor crRNA. We develop an optimized variant, denCas12m, and leverage it in the smCRISPRa platform.
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