各种疾病的分子基础PPT.ppt

各種疾病的分子基礎 單基因疾病的突變類型 單一鹼基的突變 單一鹼基的取代位置可能在 啟動子、剪接訊號（內含子改變了剪接點、表現子產生不該有的剪接點）、poly A訊號、表現子的錯義突變(missense)或無意義突變(nonsense)、終止密碼突變、起始密碼突變 任何一個鹼基都可能發生突變 靜默的突變（silence mutation） 某一核苷酸的突變並未造成胺基酸種類的改變 無意義的突變（nonsense mutation） 某一核苷酸的突變，使得轉譯時的密碼由一可以轉譯成胺基酸的密碼變成轉譯的終止密碼時稱之 錯義突變(missense) 某一核苷酸的突變，造成胺基酸序列的改變 開放讀取框架的位移突變（open reading frame shift mutation） 三個一讀核苷酸的密碼子，發生讀取位置錯位一個或兩個鹼基，使之後所轉譯的胺基酸序列都是錯誤的 黑人族群發病率1/500的鐮刀細胞貧血 隱性遺傳 Beta-globin 上的glutamic acid (GAG)變成了valine (GTG) 血紅素蛋白四聚體，低氧情況下形成不溶性 紅血球變形失去彈性 不能通過微血管 循環阻塞-引起內臟損傷 瘧疾流行區高達1/12，這可能是演化的結果 治療的契機……. Previous studies have demonstrated that sickle cell disease (SCD) can be corrected in mouse models by transduction of hematopoietic stem cells with lentiviral vectors containing antisickling globin genes followed by transplantation of these cells into syngeneic recipients. Although self-inactivating (SIN) lentiviral vectors with or without insulator elements should provide a safe and effective treatment in humans, some concerns about insertional mutagenesis persist. An ideal correction would involve replacement of the sickle globin gene (βS) with a normal copy of the gene (βA). We recently derived embryonic stem (ES) cells from a novel knock-in mouse model of SCD and tested a protocol for correcting the sickle mutation by homologous recombination. In this paper, we demonstrate the replacement of the human βS-globin gene with a human βA-globin gene and the derivation of mice from these cells. The animals produce high levels of normal human hemoglobin (HbA) and the pathology associated with SCD is corrected. Hematologic values are restored to normal levels and organ pathology is ameliorated. These experiments provide a foundation for similar studies in human ES cells derived from sickle cell patients. Although efficient methods for production of human ES cells by somatic nuclear transfer must be developed, the data in this paper demonstrate that sickle cell disease can be corrected without the risk of insertional mutagenesis. 突變熱