Multiplex Genome Editing in Chinese Hamster Ovary Cell Line Using All-in-One and HITI CRISPR Technology

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SAFARI FATEMEH; FARAJNIA SAFAR; GHASEMI YOUNES; ZARGHAMI NOSRATOLLAH; Barekati Mowahed Mazyar

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Journal Paper

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Journal: Advanced Pharmaceutical Bulletin Year:2021 | Volume:11 | Issue:2 Page(s): 343-350

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Title

Multiplex Genome Editing in Chinese Hamster Ovary Cell Line Using All-in-One and HITI CRISPR Technology

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Keywords

Apoptosis

CHO cell

Caspase 7

CRISPR-Associated Protein 9

Abstract

Purpose: CRISPR/Cas9 gene editing technology has revolutionized gene manipulation by providing the opportunity of gene knock out/in, transcriptional modification and base editing. The application of this system extended into different eras of biology, from cell development to animal modeling. Various generations of CRISPR technology have been developed to make genome editing easy which resulted in rapid protocols for amelioration of a large genome. Methods: We established a simple protocol for gene manipulation in Chinese hamster ovary (CHO) cells to achieve a Caspase 7 deficient cell line by using combination of all-in-one CRISPR technology and CRISPR/Cas9 homology-independent targeted integration (CRISPR HITI). Results: the findings of this study indicated that using CRISPR knocking in/out technology facilitates genomic manipulation in CHO cells. Integration of EGFP in target locus of Caspase 7 gene made the selection of knockout CHO cell line easy which achieved by cell sorting and single-cell cloning. Conclusion: this system introduces an effective targeting strategy for multiplex genome engineering, coinciding gene integration which simplified the selection of desired genomic characteristics.

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APA: Copy

SAFARI, FATEMEH, FARAJNIA, SAFAR, GHASEMI, YOUNES, ZARGHAMI, NOSRATOLLAH, & Barekati Mowahed, Mazyar. (2021). Multiplex Genome Editing in Chinese Hamster Ovary Cell Line Using All-in-One and HITI CRISPR Technology. ADVANCED PHARMACEUTICAL BULLETIN, 11(2), 343-350. SID. https://sid.ir/paper/784007/en

Vancouver: Copy

SAFARI FATEMEH, FARAJNIA SAFAR, GHASEMI YOUNES, ZARGHAMI NOSRATOLLAH, Barekati Mowahed Mazyar. Multiplex Genome Editing in Chinese Hamster Ovary Cell Line Using All-in-One and HITI CRISPR Technology. ADVANCED PHARMACEUTICAL BULLETIN[Internet]. 2021;11(2):343-350. Available from: https://sid.ir/paper/784007/en

IEEE: Copy

FATEMEH SAFARI, SAFAR FARAJNIA, YOUNES GHASEMI, NOSRATOLLAH ZARGHAMI, and Mazyar Barekati Mowahed, “Multiplex Genome Editing in Chinese Hamster Ovary Cell Line Using All-in-One and HITI CRISPR Technology,” ADVANCED PHARMACEUTICAL BULLETIN, vol. 11, no. 2, pp. 343–350, 2021, [Online]. Available: https://sid.ir/paper/784007/en

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