CELL LINE DEVELOPMENT THROUGH PIGGYBAC TRANSPOSON SYSTEM FOR THERAPEUTIC LYSOSOMAL ENZYMES PRODUCTION, ELOSULFASE ALFA AND IMIGLUCERASE, IN THE CHO CELL LINES.

Main Article Content

Seyed Mehdi Hassanzadeh
Azadeh Mirfeizollahi
Jafar Nikzad
Elahe Omidi
Hooman Kaghazian

Keywords

Transposon, cell line development, Elosulfase alfa, Imiglucerase, PiggyBac

Abstract

lysosomal storage diseases are treated using recombinant enzyme replacement in Morquio A syndrome by Elosulfase alfa and Gaucher disease via Imiglucerase. Production of these recombinant enzymes with high quality need to appropriate genetic manipulation of the host genome. In this work, PiggyBac transposon system was designed for expression of elosulfase alfa and imiglucerase in the Chinese hamster ovary (CHO) cells. Three transposon vectors including PB::GALNS, PB::GBA, and PB::GFP were constructed separately and together with SuperPBase vector were transfected into the CHO-K1 cells. Then recombinant cell lines were selected by exposure to zeocin. The appearance of green fluorescent cells by PB: GFP/Super Base vectors showed the efficiency of the designed transposon system. The expression of elosulfase alfa in the GALNS recombinant cell lines and imiglucerase in GBA recombinant cells were confirmed by ELISA. In conclusion, The PiggyBac transposon system can be considered as an efficient tool to modification the CHO host genome for the development of recombinant cell lines to produce therapeutic enzymes.

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