Extended Data Fig. 1: Insertion/deletion (indel) mutations generated in healthy donor CD34+ hematopoietic stem and progenitor cells (HSPCs) by Cas9 nuclease.
From: Potent and uniform fetal hemoglobin induction via base editing
a, BCL11A gene showing the target GATA1 binding motif (TGATAA) in the +58 BCL11A erythroid enhancer highlighted in green. The single-guide RNA (sgRNA) protospacer and protospacer-adjacent motifs (PAM) are shown in black and red, respectively. The vertical dotted line indicates the Cas9 cleavage site. b, The γ-globin (HBG1/2) gene showing the target BCL11A binding motif (TGACCA) in the promoter highlighted in yellow. c,d, Sequence alignments of the BCL11A and γ-globin genes showing the most common Cas9 indels determined by next-generation sequencing (NGS) 3 days after editing. The sgRNA sequences are underlined with PAM in red text. Targeted transcription factor binding motifs are indicated by the orange columns and shown in the wild type (WT) sequences as underlined blue text. Deletions are represented by dashes and insertions by green text. The % of each indel relative to total NGS reads is shown at right. e, Percentage of indels after base editing (n = 8 for UT, −198, −175 and −113, n = 6 for NT). Bar graphs show mean ± standard deviation (SD). Each symbol represents an independent experiment with different shapes representing unique HSPC donors. UT, untreated. NT, non-targeting gRNA.
