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. 2021 Jan 20;12:472. doi: 10.1038/s41467-020-20792-y

Fig. 1. Design and production of barcoded AAV6 donors for long-term genetic tracking of gene-targeted cells and their progeny.

Fig. 1

a Schematic of HBB targeting strategy. Top: Unmodified (WT) and barcoded HBB alleles depicted, with location of the E6V (GAG - > GTG) sickle cell disease mutation and CRISPR/Cas9 target sites labeled. Bottom: β-globin ORF translation with four barcode pools representing all possible silent mutations encoding amino acids 1-9. b Schematic of barcode library generation and experimental design. c/d Percentages of reads from each valid barcode identified through amplicon sequencing of plasmids (c) and AAV (d) pools 1, 2, and 4. e Recovery of barcodes from untreated genomic DNA containing 1, 3, 10, 30, and 95 individual plasmids containing HBB barcodes. Expected number of barcodes is plotted against the number of barcodes called by the TRACE-seq pipeline after filtering.