Question
is there anything about beta thalasemia yes or no The HPLC data, as clearly shown in the representative chromatograms, indicate a de
novo production of adult hemoglobin (HbA) in all GE‐samples analyzed, in agreement
with the data indicating efficient gene editing (Figures 1 and 2A,B). An increased produc‐
tion of HbF was in addition observed when the GE plus rapamycin cultures were com‐
pared with the GE‐only samples. Figure 3D,E show the summary of the treatments per‐
formed in which the GE, rapamycin and GE plus rapamycin samples were compared with
control untreated cells. The results obtained confirm that GE treatment does not interfere
with HbF induction (Figure 3D). In addition, despite the fact that with the presence of the
reactivation of γ‐globin genes the β‐globin gene expression might be analogically re‐
duced, the data obtained demonstrate that the rapamycin induction of γ‐globin genes
does not interfere with the de novo HbA production using the GE approach (Figure 3E).
The conclusions of the HPLC studies were further confirmed by the Western blotting
analyses shown in Figure 4, which aimed at evaluating the β‐globin and γ‐globin proteins
produced under the different experimental conditions depicted in Figure 3. The amounts
of CRISPR‐Cas9‐corrected β‐globin protein (16 kDa) and γ‐globin protein (15 kDa) were
normalized with the quantity of housekeeping GAPDH (37 kDa) protein. The data ob‐
tained show that rapamycin, as expected, does not induce an increase in the β‐globin pro‐
tein in control cells; in addition, rapamycin treatment does not affect the accumulation of
β‐globin in CRISPR‐Cas9 corrected ErPCs (Figure 4A).
On the other hand, the gene editing treatment has no
novo production of adult hemoglobin (HbA) in all GE‐samples analyzed, in agreement
with the data indicating efficient gene editing (Figures 1 and 2A,B). An increased produc‐
tion of HbF was in addition observed when the GE plus rapamycin cultures were com‐
pared with the GE‐only samples. Figure 3D,E show the summary of the treatments per‐
formed in which the GE, rapamycin and GE plus rapamycin samples were compared with
control untreated cells. The results obtained confirm that GE treatment does not interfere
with HbF induction (Figure 3D). In addition, despite the fact that with the presence of the
reactivation of γ‐globin genes the β‐globin gene expression might be analogically re‐
duced, the data obtained demonstrate that the rapamycin induction of γ‐globin genes
does not interfere with the de novo HbA production using the GE approach (Figure 3E).
The conclusions of the HPLC studies were further confirmed by the Western blotting
analyses shown in Figure 4, which aimed at evaluating the β‐globin and γ‐globin proteins
produced under the different experimental conditions depicted in Figure 3. The amounts
of CRISPR‐Cas9‐corrected β‐globin protein (16 kDa) and γ‐globin protein (15 kDa) were
normalized with the quantity of housekeeping GAPDH (37 kDa) protein. The data ob‐
tained show that rapamycin, as expected, does not induce an increase in the β‐globin pro‐
tein in control cells; in addition, rapamycin treatment does not affect the accumulation of
β‐globin in CRISPR‐Cas9 corrected ErPCs (Figure 4A).
On the other hand, the gene editing treatment has no
Answers
GPT 3.5
direct effect on HbF induction, and the reactivation of γ-globin genes does not interfere with de novo HbA production using the gene editing approach. The presence of rapamycin does not affect the accumulation of β-globin in CRISPR-Cas9 corrected cells. Therefore, the answer is: yes, there is information about beta thalassemia in the provided text.