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WQC, SSL, CC, and JC performed the experiments

Barcoded Read 2 FASTQs were then aligned using bwa (v0.7.17) 114 mem to a custom reference genome containing the sequence for each of the 60 gRNAs in the CROPseq library, flanked on both side by 85 bp anchor sequences from the surrounding CROPseq-RFP backbone, ensuring sufficient reference sequence to successfully align all 90 bp Read 2 sequences that overlapped at least 5 bp of the gRNA sequence