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.

通过使用人类威尔逊氏病cDNA片段，用聚合酶链反应扩增的人类威尔逊氏病cDNA片段作为探针，发现了对威尔逊氏病基因和对照wkah大鼠的限制片段长度多酰化。发现该RFLP在LEC和WKAH大鼠之间的背部交叉中显示出与肝疾病的交叉链接，表明LEC大鼠的肝疾病与人类威尔逊氏病的基因相同。该结果表明，LEC大鼠是威尔逊氏病基因疗法的使用动物模型。为了构建复古病毒载体，将人类威尔逊病基因插入了基因疗法，我们尝试从健康的人肝脏中从CDNA文库中克隆该基因cDNA。但是，获得的cDNA克隆并未编码整个编码区域，这可能是由于该基因的编码区域相对较长。我们正在重新筛选cDNA文库，以获得编码整个编码区域的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。”