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KMID : 0380020220370010017
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Korean Journal of Biotechnology and Bioengineering
2022 Volume.37 No. 1 p.17 ~ p.24
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PCR-based Analysis of Transgene Integration Sites using DNA Repeat Sequences of Recombinant CHO Cells
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Lee Sang-Yeop
Baik Jong-Youn
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Abstract
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Chinese hamster ovary (CHO) cells have been used for the production of therapeutic recombinant proteins, including monoclonal antibodies. CHO cell lines producing recombinant protein are developed traditionally by random integration, leading to clonal variation issues, arbitrarily various phenotypic characteristics between clones. Thus, development of inexpensive and fast strategy for transgene integration site analysis is necessary. In this study, we optimized Alu polymerase chain reaction (Alu PCR) for CHO cells using short interspersed nuclear elements (SINEs) in the genome. Uracilcontaining primers were used to specifically amplify the target between SINE and recombinant protein coding transgene. We applied this strategy to identify transgene integration site as well as the endogenous housekeeping gene as a control. In conclusion, we verify that Alu PCR can be an inexpensive and rapid alternative analysis tool to detect integration sites.
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KEYWORD
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Chinese hamster ovary (CHO), short interspersed nuclear elements (SINEs), Alu PCR, cell line development, transgene integration site
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