Antigen-Mediated Genetically Modified Cell Amplification (AMEGA) with Single Vector Transductio

JOURNAL OF CHEMICAL ENGINEERING OF JAPAN

Masahiro Kawahara²  Etsuji Kaneko²  Hiroshi Ueda²  Teruyuki Nagamune²  Izumi Kumagai¹  Kouhei Tsumoto¹ 

[1] Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University;Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo

关键词: gp130;    Antibody Variable Region;    Chimeric Receptor;    Transgene Selection;    Gene Transduction;   

DOI  :  10.1252/jcej.37.1259

来源: Maruzen Company Ltd

【 摘 要 】

References(18)Cited-By(3)Previously, we proposed the Antigen-mediated genetically modified cell amplification (AMEGA) system, which can selectively amplify gene-transduced cells without antibiotic selection. While the original AMEGA system employed two vectors encoding a pair of antibody/receptor chimeras and a gene of interest, the use of two virions resulted in low co-transduction efficiency and time-consuming selection. Here we show an improved AMEGA system, where the three genes are linked in tandem with internal ribosomal entry sites in a single vector. We used gp130 chimeras whose extracellular domain was replaced with either VH or VL region of anti-hen egg lysozyme (HEL) antibody HyHEL-10 and D2 domain of erythropoietin receptor. The constructed vector was retrovirally transduced to IL-3-dependent Ba/F3 cells followed by HEL selection in the absence of IL-3. The resultant transduction efficiency increased by ~100-fold compared to the previous two-vector system, which results in a much shorter selection period.

【 授权许可】

Unknown