American Society of Hematology

Figure 1.

$A protein domain–focused CRISPR screen identified BMI1 as a novel HbF repressor. (A) Scatter plot of sgRNA abundance in HbF-high and HbF-low populations from the domain-focused CRISPR screen. Each dot represents 1 sgRNA. Control sgRNAs (n = 50) and sgRNAs targeting BMI1 (n = 6) are labeled in blue and red, respectively. (B-D) Representative immunoblots of BMI1, GATA1, and γ-globin protein, HBG:(HBG+HBB) mRNA level, and HbF+ cell fraction in control (sgNeg: nontargeting sgRNA; sgBCL11A+58: positive control) and BMI1-depleted HUDEP2 cells. n = 2. β-Actin was used as the loading control in immunoblot experiment. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as internal control for RT-qPCR. Values are presented as mean ± SEM. SEM, standard error of the mean.$

A protein domain–focused CRISPR screen identified BMI1 as a novel HbF repressor. (A) Scatter plot of sgRNA abundance in HbF-high and HbF-low populations from the domain-focused CRISPR screen. Each dot represents 1 sgRNA. Control sgRNAs (n = 50) and sgRNAs targeting BMI1 (n = 6) are labeled in blue and red, respectively. (B-D) Representative immunoblots of BMI1, GATA1, and γ-globin protein, HBG:(HBG+HBB) mRNA level, and HbF⁺ cell fraction in control (sgNeg: nontargeting sgRNA; sgBCL11A+58: positive control) and BMI1-depleted HUDEP2 cells. n = 2. β-Actin was used as the loading control in immunoblot experiment. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as internal control for RT-qPCR. Values are presented as mean ± SEM. SEM, standard error of the mean.

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