Enhanced Transgene Expression by Optimization of Poly A in Transfected CHO Cells. Wang, X., Du, Q., Zhang, W., Xu, D., Zhang, X., Jia, Y., & Wang, T. Frontiers in Bioengineering and Biotechnology, jan, 2022. Paper doi abstract bibtex The generation of the stable, high-level recombinant protein-producing cell lines remains a significant challenge in the biopharmaceutical industry. Expression vector optimization is an effective strategy to increase transgene expression levels and stability, and the choice of suitable poly A element is crucial for the expression of recombinant protein. In this study, we investigated the effects of different poly A elements on transgene expression in Chinese hamster ovary (CHO) cells. Five poly A elements, including bovine growth hormone (BGH), mutant BGH, herpes simplex virus type 1 thymidine kinase (HSV-TK), SV40, and a synthetic (Synt) poly A, were cloned into the expression vector and transfected into CHO cells. The results indicated the SV40 and Synt poly A sequences can significant improve eGFP transgene expression in stable transfected CHO cells and maintain long-term expression. However, qPCR results showed that the eGFP expression at protein level was not related to the gene copy number and mRNA level. Importantly, the SV40 and Synt poly A elements decreased the variation of eGFP transgene expression. Furthermore, it also showed that the SV40 and Synt poly A elements induced higher levels of adalimumab expression. In conclusion, SV40 poly A and Synt poly A are stronger elements that increase stable transgene expression and decrease the variation of expression, and the choice of suitable poly A element is helpful to improve the expression of recombinant protein.
@article{Wang2022,
abstract = {The generation of the stable, high-level recombinant protein-producing cell lines remains a significant challenge in the biopharmaceutical industry. Expression vector optimization is an effective strategy to increase transgene expression levels and stability, and the choice of suitable poly A element is crucial for the expression of recombinant protein. In this study, we investigated the effects of different poly A elements on transgene expression in Chinese hamster ovary (CHO) cells. Five poly A elements, including bovine growth hormone (BGH), mutant BGH, herpes simplex virus type 1 thymidine kinase (HSV-TK), SV40, and a synthetic (Synt) poly A, were cloned into the expression vector and transfected into CHO cells. The results indicated the SV40 and Synt poly A sequences can significant improve eGFP transgene expression in stable transfected CHO cells and maintain long-term expression. However, qPCR results showed that the eGFP expression at protein level was not related to the gene copy number and mRNA level. Importantly, the SV40 and Synt poly A elements decreased the variation of eGFP transgene expression. Furthermore, it also showed that the SV40 and Synt poly A elements induced higher levels of adalimumab expression. In conclusion, SV40 poly A and Synt poly A are stronger elements that increase stable transgene expression and decrease the variation of expression, and the choice of suitable poly A element is helpful to improve the expression of recombinant protein.},
author = {Wang, Xiao-yin and Du, Qiu-jie and Zhang, Wei-li and Xu, Dan-hua and Zhang, Xi and Jia, Yan-long and Wang, Tian-yun},
doi = {10.3389/fbioe.2022.722722},
issn = {2296-4185},
journal = {Frontiers in Bioengineering and Biotechnology},
keywords = {Chinese hamster ovary cells,optimization,poly A elements,stability,transgene expression},
month = {jan},
title = {{Enhanced Transgene Expression by Optimization of Poly A in Transfected CHO Cells}},
url = {https://www.frontiersin.org/articles/10.3389/fbioe.2022.722722/full},
volume = {10},
year = {2022}
}
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Five poly A elements, including bovine growth hormone (BGH), mutant BGH, herpes simplex virus type 1 thymidine kinase (HSV-TK), SV40, and a synthetic (Synt) poly A, were cloned into the expression vector and transfected into CHO cells. The results indicated the SV40 and Synt poly A sequences can significant improve eGFP transgene expression in stable transfected CHO cells and maintain long-term expression. However, qPCR results showed that the eGFP expression at protein level was not related to the gene copy number and mRNA level. Importantly, the SV40 and Synt poly A elements decreased the variation of eGFP transgene expression. Furthermore, it also showed that the SV40 and Synt poly A elements induced higher levels of adalimumab expression. In conclusion, SV40 poly A and Synt poly A are stronger elements that increase stable transgene expression and decrease the variation of expression, and the choice of suitable poly A element is helpful to improve the expression of recombinant protein.","author":[{"propositions":[],"lastnames":["Wang"],"firstnames":["Xiao-yin"],"suffixes":[]},{"propositions":[],"lastnames":["Du"],"firstnames":["Qiu-jie"],"suffixes":[]},{"propositions":[],"lastnames":["Zhang"],"firstnames":["Wei-li"],"suffixes":[]},{"propositions":[],"lastnames":["Xu"],"firstnames":["Dan-hua"],"suffixes":[]},{"propositions":[],"lastnames":["Zhang"],"firstnames":["Xi"],"suffixes":[]},{"propositions":[],"lastnames":["Jia"],"firstnames":["Yan-long"],"suffixes":[]},{"propositions":[],"lastnames":["Wang"],"firstnames":["Tian-yun"],"suffixes":[]}],"doi":"10.3389/fbioe.2022.722722","issn":"2296-4185","journal":"Frontiers in Bioengineering and Biotechnology","keywords":"Chinese hamster ovary cells,optimization,poly A elements,stability,transgene expression","month":"jan","title":"Enhanced Transgene Expression by Optimization of Poly A in Transfected CHO Cells","url":"https://www.frontiersin.org/articles/10.3389/fbioe.2022.722722/full","volume":"10","year":"2022","bibtex":"@article{Wang2022,\nabstract = {The generation of the stable, high-level recombinant protein-producing cell lines remains a significant challenge in the biopharmaceutical industry. Expression vector optimization is an effective strategy to increase transgene expression levels and stability, and the choice of suitable poly A element is crucial for the expression of recombinant protein. In this study, we investigated the effects of different poly A elements on transgene expression in Chinese hamster ovary (CHO) cells. Five poly A elements, including bovine growth hormone (BGH), mutant BGH, herpes simplex virus type 1 thymidine kinase (HSV-TK), SV40, and a synthetic (Synt) poly A, were cloned into the expression vector and transfected into CHO cells. The results indicated the SV40 and Synt poly A sequences can significant improve eGFP transgene expression in stable transfected CHO cells and maintain long-term expression. However, qPCR results showed that the eGFP expression at protein level was not related to the gene copy number and mRNA level. Importantly, the SV40 and Synt poly A elements decreased the variation of eGFP transgene expression. 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