Basic research for the Wilson's disease gene therapy

威尔逊病基因治疗的基础研究

• 批准号: 06454304
• 负责人: WADA Yoshiro
• 金额: $ 4.61万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 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/en/grant/KAKENHI-PROJECT-06454304/
• 关键词: Wilson's desease; Gene therapy; LEC rats

Restriction fragment length polymoyphism on the Wilson's disease gene betweem LEC and control WKAH rats was found by using a human Wilson's disease cDNA fragment amplified with polymerase chain reaction as the probe. This RFLP was found to show a cross-linkage with the hepatic disorder in back-crosses between LEC and WKAH rats, indicating that the hepatic disorder in LEC rats is due to the same gene as the case of human Wilson's disease. This result suggests that LEC rats are usuful animal model for the Wilson's disease gene therapy.In order to construct a retro virus vector inserted the human Wilson's disease gene for the gene therapy, we tried cloning of this gene cDNA from the cDNA library made from the healthy human liver. However, cDNA clones obtained all did not encode the whole coding region, possibly due to the relatively long coding region of this gene. We are under re-screening the cDNA library to obtain cDNA encoding the whole coding region.

以聚合酶链式反应扩增的人威尔逊氏病cDNA片段为探针，发现LEC和对照WKAH大鼠威尔逊氏病基因的限制性片段长度多态性。在 LEC 和 WKAH 大鼠回交中发现该 RFLP 显示出与肝脏疾病的交联，表明 LEC 大鼠的肝脏疾病与人类威尔逊氏病的情况是由相同的基因引起的。这一结果表明LEC大鼠是威尔逊氏病基因治疗的有用动物模型。为了构建插入人类威尔逊氏病基因用于基因治疗的逆转录病毒载体，我们尝试从健康人肝脏制备的cDNA文库中克隆该基因cDNA。然而，获得的cDNA克隆均不编码整个编码区，可能是由于该基因的编码区相对较长。我们正在重新筛选cDNA文库以获得编码整个编码区的cDNA。

项目成果

N.Sasaki et al.: "The gene responsible for LEC hepatitis,licated on rat chromosome 16,is the homolog to the human Wilson disease gene" Biochem.Biophys.Res.Commun.202. 512-518 (1994)

N.Sasaki 等人：“导致 LEC 肝炎的基因位于大鼠 16 号染色体上，与人类威尔逊病基因同源”Biochem.Biophys.Res.Commun.202。

N. Sasaki et al.: "The gene responsible for LEC hepatitis,located on rat chromosome 16,is the homolog to the human Wilson disease gene." Biochem. Biophys. Res. Commun.202. 512-518 (1994)

N. Sasaki 等人：“导致 LEC 肝炎的基因位于大鼠 16 号染色体上，与人类威尔逊病基因同源。”

N.Sasaki et al.: "The gene responsible for LEC hepatitis, located on rat chromosome 16, is the homolog to the human Wilson disease gene." Biochem.Biophys.Res.Commun.202. 512-518 (1994)

N.Sasaki 等人：“导致 LEC 肝炎的基因位于大鼠 16 号染色体上，与人类威尔逊病基因同源。”