چاپ صفحه |  صفحه نخست سامانه |  چکیده مقاله |  نویسندگان |  دانلود مقاله | دانشگاه علوم پزشکی تبریز |
| Background: Chinese hamster ovary (CHO) cells are the most commonly used mammalian host cell in the commer‑ cial‑scale production of biopharmaceutical proteins. Modification of genes involved in apoptosis may improve the productivity of CHO cells. Executive caspases, including caspases 3 and 7, play critical roles in apoptosis. The effects of the ablation of the caspase 7 gene on proliferation and viability of CHO cells remains unknown. In this study, we applied clustered regularly interspaced short palindromic repeat (CRISPR/Cas9) to target caspase 7 gene of CHO K1 cell via all in one and homology targeted integration strategies. Consequently, the effect of caspase 7 deficiency on cell proliferation, viability, and apoptosis was studied by MTT assay and flow cytometry. Results: Findings of gel electrophoresis, western blotting, and sequencing confirmed the caspase 7 gene silencing in CHO cells (CHO‑KO). Proliferation assay revealed that caspase 7 deficiency in CHO cells resulted in the reduction of proliferation in various CHO‑KO clones. Besides, the disruption of caspase 7 had negative effects on cell viability in exposure with NaBu which confirmed by MTT assay. Results of flow cytometry using Anexin V/PI demonstrated that Nabu treatment (11 mM) declined the percentage of live CHO‑K1 and CHO‑KO cells to 70.3% and 5.79%. These results verified that the CHO‑K1 cells were more resistant to apoptosis than CHO‑KO, however most of CHO‑KO cells under‑ gone early apoptosis (91.9%) which seems to be a fascinating finding. Conclusion: These results reveal that caspase 7 may be involved in the cell cycle progression of CHO cells. Further‑ more, it seems that targeting caspase 7 is not the ideal route as it had previously been imagined within the preven‑ tion of apoptosis but the relation between caspase 7 deficiency, cell cycle arrest, and the occurrence of early apopto‑ sis will require more investigation |
| نویسنده | نفر چندم مقاله |
|---|---|
| فاطمه صفری | اول |
| صفر فرج نیا | دوم |
| حبیب ذره دار | چهارم |
| نام فایل | تاریخ درج فایل | اندازه فایل | دانلود |
|---|---|---|---|
| cas-7.pdf | 1399/11/28 | 1933139 | دانلود |
