Based on the data presented in the figure, it appears that the combination of CRISPR-Cas9 gene editing and rapamycin treatment has a positive effect on the expression of β-globin mRNA in ErPCs isolated from β039-thalassemia patients. This combination treatment did not have a significant impact on the expression of γ-globin mRNA, indicating that CRISPR-Cas9 gene editing does not interfere with rapamycin-mediated induction of γ-globin gene expression. Additionally, there was no change in the expression of α-globin mRNA, suggesting that the treatment did not affect the expression of α-globin genes in ErPCs.
Overall, these findings suggest that the combination of CRISPR-Cas9 gene editing and rapamycin treatment could potentially be a promising approach for treating β-thalassemia by enhancing the expression of β-globin genes. However, further research and clinical studies would be needed to determine the efficacy and safety of this treatment approach in patients with thalassemia.
is there aything about using cas09 to treat thalasmia
Figure 2. Evaluation of the combination between CRISPR‐Cas9 gene editing and rapamycin‐medi‐
ated HbF induction. (A,B) Fractional abundance obtained after treatment performed by CRISPR‐
Cas9 system on ErPCs isolated from β039‐thalassemia patients and analyzed by ddPCR assay. The
histograms show the data related to β‐globin gene (A) and β‐globin mRNA (B). (C–E) The histo‐
grams show the relative content of the β‐, γ‐ and α‐globin mRNAs analyzed by multiplex RT‐qPCR.
(−): untreated cells; GE: cells treated with the CRISPR‐Cas9 system; RAPA: rapamycin 200 nM
treated cells; GE + RAPA: cells treated with the CRISPR‐Cas9 system and cultured in the presence
of rapamycin. All the data of RT‐qPCR were normalized using GAPDH as housekeeping internal
control gene, as described in Material and Methods. Results are expressed as fold changes with re‐
spect to control untreated cells (−). Results are from independent experiments using ErPCs cultures
from three (A) and five (B–E) homozygous β039‐thalassemia patients. The level of statistical signifi‐
cance is reported as p‐value (p).
Moreover, when rapamycin and GE plus rapamycin cultures were compared, no sig‐
nificant change in the accumulation of γ‐globin mRNA was observed (p = 0.5176), demon‐
strating that gene editing has no major effects on the rapamycin‐mediated induction of
the expression of γ‐globin genes. Interestingly, no change in α‐globin mRNA content was
found in the ErPCs populations, indicating that the expression of α‐globin genes in ErPCs
treated with GE, rapamycin and GE plus rapamycin is similar to control untreated ErPCs
(−). The data shown in Figure 2C–E were obtained using GAPDH sequences as internal
reference. However, the same conclusion can be reached using RPL13A or β‐actin internal
controls (unpublished results).
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