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Scalable biological signal recording in mammalian cells using Cas12a base editors

Nat Chem Biol. 2022-05; 
Hannah R Kempton, Kasey S Love, Lucie Y Guo, Lei S Qi
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Proteins, Expression, Isolation and Analysis … Secreted antiTNF nanobody was measured using the His Tag ELISA Detection Kit (Genscript), with all samples run in technical duplicate on the ELISA plate. Absorbance was measured … Get A Quote

摘要

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 chimeric antig... More

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