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The crRNA library was delivered into cells stably expressing Cas12a by lentiviral infection. Infected cells were collected by fluorescence-activated cell sorting (FACS; green fluorescence). For screening, cells were cultured for 15-45 d before genome DNA extraction and high-throughput sequencing analysis of the barcoded crRNA regions. Each DNA oligonucleotide sequence encodes two crRNAs (represented in red and blue), which will be transcribed and processed to generate individual mature crRNAs by Cas12a; these mature crRNAs will guide Cas12a to cut target genome regions. DR (19 nt).

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