재조합 CHO 세포주의 DNA 반복서열을 활용한 PCR 기반 외래유전자 삽입위치 분석

초록

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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