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. 2021 May 1;29(9):2841–2853. doi: 10.1016/j.ymthe.2021.04.037

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© 2021 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Analysis of LVβ-shα2-mediated knockdown of α2-globin in primary erythroid cells

(A) Erythroid differentiation profile of cells from P1 grown in medium 1 after transduction with BB305, LVβ-shSCR, or LVβ-shα2 at equivalent MOIs. (B) Mean percentages of populations 1 (CD71^hi/CD36^hi), 2 (CD71^hi/CD36^med), and 3 (CD71^med/CD36^lo) in GPA⁺ cells. (C) Analysis of α/(β+γ)-globin mRNA ratios in CB and P1 cells transduced with the BB305, LVβ-shSCR, and LVβ-shα2 vectors. Results from BB305 and LVβ-shSCR were pooled. (D) VCNs in the samples analyzed in (C). Data are presented as mean +/- SD of three replicates (E) Analysis of α2/α1-globin mRNA ratios in cells from 2 individuals with HbE/β-thalassemia (P1 and P2). For each analyzed sample, ratios were normalized to those measured at the same time point in non-transduced cells. (F) Correlation between α2/β-globin and α1/β-globin mRNA ratios in erythroid cells of P1 (days 11, 14, and 16) and P2 (days 11, 15, 18, and 21).