MAR elements as tools to increase protein production by CHO cells

Détails

ID Serval
serval:BIB_4C16B97533D7
Type
Actes de conférence (partie): contribution originale à la littérature scientifique, publiée à l'occasion de conférences scientifiques, dans un ouvrage de compte-rendu (proceedings), ou dans l'édition spéciale d'un journal reconnu (conference proceedings).
Collection
Publications
Titre
MAR elements as tools to increase protein production by CHO cells
Titre de la conférence
Animal Cell Technology Meets Genomics.
Auteur(s)
Girod P.A., Zahn-Zabal M.M., Mermod N.
Editeur
Springer
Organisation
Proceedings of the 18th ESACT Meeting Granada, Spain, May 11-14, 2003
Adresse
Dordrecht
ISBN
978-1-4020-2791-8
Statut éditorial
Publié
Date de publication
2005
Editeur scientifique
Gòdia F., Fussenegger M.
Volume
2
Série
ESACT Proceedings
Pages
411-415
Langue
anglais
Résumé
One of the major hurdles of isolating stable, inducible or constitutive high-level producer cell lines is the time-consuming selection, analysis and characterization of the numerous clones required to identify one with the desired characteristics. Various boundary elements, matrix attachment regions, and locus control regions were screened for for their ability to augment the expression of heterologous genes in CHO and other cells. The 5'-matrix-attachment region (MAR) of the chicken lysozyme gene was found to significantly increase stable gene expression, in culture dishes and in bioreactors. These MAR elements can be easily combined with various existing expression systems, as they can be added in trans (i.e. on a separate plasmid) in co-transfections with previously constructed expression vectors. Using cell population analysis, we found that the use of the MAR increases the proportion of high-producing CHO cell clones, thus reducing the number of cell lines that need to be screened while increasing maximal productivity. Random cDNA cloning and sequencing indicated that over 12% of the ESTs correspond to the transgene. Thus, productivity is no longer limited by transcriptional events in such MAR-containing cell lines. The identification of small and more convenient active MAR portions will also be summarized. Finally, we will show examples of how MAR elements can be combined with short term expression to increase the simultaneous synthesis of many proteins in parallel by CHO cells. Overall, we conclude that the MAR sequence is a versatile tool to increase protein expression in short and long term production processes.
Création de la notice
16/06/2010 11:09
Dernière modification de la notice
20/08/2019 14:00
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