Highlights d Spontaneously dimerizing split Cas12a for construction of complex genetic circuits d Implementation of robust 2-, 3-, and 4-input AND gates to control endogenous genes d Split Cas12a, inducible guides, and anti-CRISPR allow higher order logic computation d Circuits can detect tumor-relevant signals for therapeutic gene expression

No abstract

Biological signal recording enables the study of molecular inputs experienced throughout cellular history. However, current methods are limited in their ability to scale up beyond a single signal in mammalian contexts. Here, we develop an approach using a hyper-efficient dCas12a base editor for multi-signal parallel recording in human cells. We link signals of interest to expression of guide RNAs to catalyze specific nucleotide conversions as a permanent record, enabled by Cas12’s guide-processing abilities. We show this approach is plug-and-play with diverse biologically relevant inputs and extend it for more sophisticated applications, including recording of time-delimited events and history of CAR-T cells’ antigen exposure. We also demonstrate efficient recording of up to four signals in parallel on an endogenous safe-harbor locus. This work provides a versatile platform for scalable recording of signals of interest for a variety of biological applications.