Generating a transgenic mouse line stably expressing human MHC surface antigen from a HAC carrying multiple genomic BACs.

Generating a transgenic mouse line stably expressing human MHC surface antigen from a HAC carrying multiple genomic BACs.
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DOI:
10.1007/s00412-014-0488-3
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发表时间:
2015-03
期刊:
影响因子:
1.6
通讯作者:
Masumoto H
Masumoto H
中科院分区:
生物学3区
文献类型:
--
作者:
Hasegawa Y;Ishikura T;Hasegawa T;Watanabe T;Suzuki J;Nakayama M;Okamura Y;Okazaki T;Koseki H;Ohara O;Ikeno M;Masumoto H

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人类人工染色体(HAC)载体是一种很有前途的工具,可以改善传统质粒或病毒载体产生的转基因细胞和动物中常见的转基因表达抑制和定位效应问题。我们生成了转基因小鼠,该小鼠维持单个 HAC 载体,该载体携带来自人类 HLA-DR 基因座(DRA 和 DRB1)的两条基因组细菌人工染色体(BAC)。 RT-PCR 分析显示,转基因小鼠 HAC 上的两种转基因在肾、肝、肺、脾、淋巴结、骨髓和胸腺细胞中均表现出组织特异性表达。通过脾细胞的流式细胞术分析检测到 HAC 上两个转基因 DRA 和 DRB1 的细胞表面 HLA-DR 标记的稳定功能表达,并维持至少八个子代。这些结果表明,从头HAC系统可以让我们通过产生转基因动物来操纵多个BAC转基因,并作为表面抗原协调表达。本文的在线版本 (doi:10.1007/s00412-014-0488-3) 包含补充材料,可供授权用户使用。
The human artificial chromosome (HAC) vector is a promising tool to improve the problematic suppression and position effects of transgene expression frequently seen in transgenic cells and animals produced by conventional plasmid or viral vectors. We generated transgenic mice maintaining a single HAC vector carrying two genomic bacterial artificial chromosomes (BACs) from human HLA-DR loci (DRA and DRB1). Both transgenes on the HAC in transgenic mice exhibited tissue-specific expression in kidney, liver, lung, spleen, lymph node, bone marrow, and thymus cells in RT-PCR analysis. Stable functional expression of a cell surface HLA-DR marker from both transgenes, DRA and DRB1 on the HAC, was detected by flow cytometric analysis of splenocytes and maintained through at least eight filial generations. These results indicate that the de novo HAC system can allow us to manipulate multiple BAC transgenes with coordinated expression as a surface antigen through the generation of transgenic animals. The online version of this article (doi:10.1007/s00412-014-0488-3) contains supplementary material, which is available to authorized users.
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