Transgenic analysis of the expression of OPN gene-encoding a molecule responsible for cancer cell attachment.

对编码负责癌细胞附着的分子的 OPN 基因表达进行转基因分析。

• 批准号: 06044073
• 负责人: NODA Masaki
• 金额: $ 5.44万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06044073/
• 关键词: Osteopontin; Osteoblat; Bone Morphogenetic Protein; Bone Formation; Bone Protein; Osteoclast; gene expression; Differentiation; osteopontin; osteoblat; osteoclast

To eluidate the role of osteopontinm, we prepared to examine the effect of osteopontin overexpression as well as inactivation of osteopontin gene in mice system. We first cloned murine osteopontin gene and constructed vectors by using the upstream exons. These constructs also contained neo-gene cassett and were introduced into murine ES cells by electroporation. ES cells containing homologous recombination in the osteopontin gene were selected and they were introduced into the blastocele. Chymeric mice were identified by their coat color and were mated to prodece mice which lack osteopontin gene through homolo gous recombination. With regard to over expression experiments, osteopontin gene was cloned down stream of a strong promoter to yield inducible expression vectors and this vector was introduced via microinjection into fertilized eggs which were subsequently implanted into the uterus of pseudo-pregnant mice.

为了阐明骨桥蛋白的作用，我们准备检查骨桥蛋白过度表达以及骨桥蛋白基因失活对小鼠系统的影响。我们首先克隆了小鼠骨桥蛋白基因，并利用上游外显子构建了载体。这些构建体还含有新基因盒，并通过电穿孔引入小鼠 ES 细胞中。选择骨桥蛋白基因中含有同源重组的ES细胞并将其引入囊胚。通过毛色鉴定嵌合体小鼠，并通过同源重组与缺乏骨桥蛋白基因的小鼠交配。关于过表达实验，将骨桥蛋白基因克隆到强启动子下游以产生诱导型表达载体，并通过显微注射将该载体引入受精卵中，随后将受精卵植入假怀孕小鼠的子宫中。

项目成果

Hiroko N.Matsumoto: "Establishment and Characterization of Bone Marrow Stromal Cell Line That Support Osteoclastogenesis." Endocrinology. 136(9). 4084-4091 (1995)

Hiroko N.Matsumoto：“支持破骨细胞生成的骨髓基质细胞系的建立和表征。”

Malyankar,U.: "The MRP/Proliferin transcript is deg raded in primary mouse embryo fibroblost but not 3T3 nuclei:alfered RNA processing cottelates with immortalization" Proceeding National Academy of Science U.S.A. 91. 335-339 (1994)

Malyankar，U.：“MRP/Proliferin 转录物在原代小鼠胚胎成纤维细胞中被降解，但在 3T3 细胞核中未被降解：产生的 RNA 加工与永生化”，Proceeding National Academy of Science U.S.A. 91. 335-339 (1994)

Masato Tamura: "1 α,25-dihydroxyvitamin D down-regulates pleiotrophin messenger RNA expression in osteoblast-like cells." Endocrine. 3. 21-24 (1995)

Masato Tamura：“1 α,25-二羟基维生素 D 下调成骨细胞样细胞中的多效素信使 RNA 表达。”内分泌。

Lopez,C.A.: "Activation of a signal transduction pathway by Osteopontin. Osteopontin : Role in Cell Signalling and Adhesion Annals of the NY Academy of Sciences 760" The New York Academy of Sciences, 388(324-326) (1995)

Lopez,C.A.：“骨桥蛋白激活信号转导途径。骨桥蛋白：在细胞信号传导和粘附纽约科学院年鉴 760 中的作用”纽约科学院，388(324-326) (1995)

David, T.Denhardt: The New York Academy of Sciences. Structure-function atudies of mouse tissue inhibitor of metalloproteinase-1. Inhibition of Matrix Metalloproteinases : Therapeutic Potential. Annals of the NY Academy of Sciences 732, 65-74 (1994)

David, T.Denhardt：纽约科学院。