Multiplex Genome Editing in Chinese Hamster Ovary Cell Line Using All-in-One and HITI CRISPR Technology.
| Autor: | Safari F; Medical Biotechnology Department, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.; Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran., Farajnia S; Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran., Ghasemi Y; Department of Pharmaceutical Biotechnology, School of Pharmacy, and Pharmaceutical Sciences Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran., Zarghami N; Medical Biotechnology Department, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran., Barekati Mowahed M; Department of Physiology & Biophysics, School of Medicine, Case Western Reserve University, Ohio, USA. |
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| Jazyk: | angličtina |
| Zdroj: | Advanced pharmaceutical bulletin [Adv Pharm Bull] 2021 Feb; Vol. 11 (2), pp. 343-350. Date of Electronic Publication: 2020 Apr 15. |
| DOI: | 10.34172/apb.2021.032 |
| Abstrakt: | 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. (© 2021 The Authors.) |
| Databáze: | MEDLINE |
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