Asimakopoulos Fotis, Varmus Harold E
Department of Medicine, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA.
J Virol. 2009 May;83(10):4835-43. doi: 10.1128/JVI.02254-08. Epub 2009 Mar 11.
The transcription factor Blimp-1 has emerged as a regulator of cell fate in embryonic (germ cell) and adult (B- and T-cell immune effector and epithelial) lineages. It has also been proposed to act as a tumor suppressor in B-cell malignancy. Here, we present a novel in vivo system enabling the targeted genetic manipulation of cells expressing Prdm1, the gene encoding Blimp-1. We created bacterial artificial chromosome-transgenic mice expressing the avian leukosis virus (ALV) receptor TVB, fused to monomeric red fluorescent protein, under regulation by Prdm1 transcriptional elements, and we achieved transduction of TVB-expressing lymphocytes by ALV vectors bearing a subgroup B envelope. The system presented here incorporates a number of innovations. First, it is the first mammalian transgenic system that employs the ALV receptor TVB, thus expanding the flexibility and scope of ALV-mediated gene delivery. Second, it represents the first ALV-based system that allows gene transfer and expression into in vivo-activated mature lymphocytes, a cell type that has traditionally presented formidable challenges to efficient retroviral transduction. Third, Prdm1:TVB-mRFP transgenic animals could provide an invaluable tool for exploring the diverse roles of Blimp-1 in lineage commitment, immune regulation, and tumorigenesis.
转录因子Blimp-1已成为胚胎(生殖细胞)和成体(B细胞和T细胞免疫效应细胞及上皮细胞)谱系中细胞命运的调节因子。它还被认为在B细胞恶性肿瘤中发挥肿瘤抑制作用。在此,我们展示了一种新型的体内系统,该系统能够对表达Prdm1(编码Blimp-1的基因)的细胞进行靶向基因操作。我们构建了细菌人工染色体转基因小鼠,其在Prdm1转录元件的调控下表达与单体红色荧光蛋白融合的禽白血病病毒(ALV)受体TVB,并且我们通过携带B亚群包膜的ALV载体实现了对表达TVB的淋巴细胞的转导。这里展示的系统包含多项创新。首先,它是首个采用ALV受体TVB的哺乳动物转基因系统,从而扩展了ALV介导的基因递送的灵活性和范围。其次,它是首个基于ALV的系统,能够将基因转移并表达至体内活化的成熟淋巴细胞中,而这种细胞类型传统上一直是高效逆转录病毒转导面临巨大挑战的细胞类型。第三,Prdm1:TVB-mRFP转基因动物可为探索Blimp-1在谱系定向、免疫调节和肿瘤发生中的多种作用提供宝贵工具。