Selection of genetically modified cell population using hapten-specific antibody/receptor chimera
Selection of genetically modified cell population using hapten-specific antibody/receptor chimera
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DOI：
10.1016/j.bbrc.2004.01.030
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发表时间：
2004-02-27
影响因子：
3.1
通讯作者：
Nagamune, T
中科院分区：
文献类型：
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作者：
Kawahara, M;Kimura, H;Nagamune, T
对转基因细胞群体的有效选择是获得具有所需特性的细胞的关键步骤。在这项研究中，我们提出了一种抗原介导的遗传修饰细胞扩增系统(AMEGA)，该系统使用抗体/受体嵌合体来触发生长信号，以响应无毒的半抗原二聚体。通过逆转录病毒感染依赖IL-3的BA/F3细胞，将融合到促红细胞生成素受体胞外D2结构域和gp130跨膜/胞内区的抗荧光素单链抗体与IRES序列下游的模型转基因增强型绿色荧光蛋白(EGFP)一起表达。加入由不同寡核苷酸连接的荧光素二聚体可以诱导转染体的选择性生长，从而导致EGFP阳性细胞群的有效扩张。此外，特定的限制性内切酶对寡核苷酸的消化完全抑制了细胞的生长。由于这些半抗原二聚体对正常细胞无害，因此该方法对于用于细胞治疗和组织工程的转基因细胞群体的体外或体内可逆扩增将特别有用。(C)2004 Elsevier Inc.保留所有权利。
Efficient selection of the genetically modified cell population is a critical step to obtain the cells with desired properties. In this study, we propose an antigen-mediated genetically modified cell amplification (AMEGA) system employing an antibody/receptor chimera that triggers a growth signal in response to a non-toxic hapten dimer. An anti-fluorescein single-chain Fv fused to the extracellular D2 domain of erythropoietin receptor and transmembrane/intracellular domains of gp130 was expressed together with a model transgene, enhanced green fluorescent protein (EGFP) downstream of IRES sequence, by retroviral infection to IL-3-dependent Ba/F3 cells. Addition of fluorescein dimers connected by various oligo-DNA linkers induced selective growth of transfectants, thus leading to efficient expansion of EGFP-positive cell population. Also, digestion of the oligonucleotides by specific restriction endonuclease completely suppressed cell growth. Because these hapten dimers are not harmful for normal cells, the approach will be especially useful for reversible in vitro or in vivo expansion of genetically modified cell population employed for cell therapy and tissue engineering. (C) 2004 Elsevier Inc. All rights reserved.