中文摘要：

基因治疗提供潜在的痊愈因为出血不止 B，和重要进步在指导肝的基因转移被完成了由联系 adeno 的病毒的向量调停了。包含优化 codon 的因素 IX (AAV8-hFIXco ) 向量表明了鼓励功效， hFIX 表示在 1% ～ 6% 正常稳定了的自我补足的联系 adeno 的病毒 serotype 8 人的使用的最近的临床的试用在与高向量剂量有关的病人，而是安全担心铺平仍然是现在。因此， AAV 向量的进一步的改进和 hFIX 表示盒子可以断然贡献出血不止 B 基因治疗的最终的成功。在这研究，加强的 hFIX 铺平接受者的凝结剂能力获得更高的表情，人的改正表示向量被升级 codon 改编索引并且调整 GC 内容优化，插入一个 Kozak 序列( GCCACC )，并且介绍一个 gain-of-function 变化， R338L (改正 Padua )。效率屏蔽出版，目前构造的盒子直在 vivo 被比较。另外，在这些盒子控制改正基因表示的规章的元素为肝特定的有效性被屏蔽。在所有构造盒子之中， scAAV-Pre-hFIXco-SIH-R338L ，是在凝血素 enhancer 和 prealbumin 倡导者的控制下面的构造，导致了凝结剂活动的高水平，并且二个构造盒子( scAAV-Chi-hFIXco-SIH-R338L 和 scAAV-Pre-hFIXco-SIH-R338L )的表示层次也比出版盒子( scAAV-LP1-hFIXco-SJ )的高。在摘要，我们的策略在蛋白质水平或一项显著地提高的凝结剂活动在 hFIX 表示导致了实质的增加。因此，这些与 AAV hFIX 重建向量可以潜在地贡献出血不止 B 的有效基因治疗的创造。

英文摘要：

Gene therapy provides a potential cure for hemophilia B, and significant progress has been achieved in liver-directed gene transfer mediated by adeno-associated viral vectors. Recent clinical trials involving the use of a self-complementary adeno-associated virus serotype 8-human codon-optimized factor IX （AAV8-hFIXco） vector demonstrated encouraging efficacy with hFIX expression stabilized at 1% to 6% of normal level in patients, but safety concerns related to high vector doses are still present. Thus, further improvement of AAV vectors and hFIX expression cassette may positively contribute to the ultimate success of hemophilia B gene therapy. In this study, to obtain a higher expression level of hFIX that potentiates the coagulant capacity of recipients, human FIX expression vector was optimized by upgrading the codon adaption index and adjusting the GC content, inserting a Kozak sequence （GCCACC）, and introducing a gain-of-function mutation, R338L （FIX Padua）. The efficiency of the published and the presently constructed cassettes was compared through in vivo screening. In addition, the regulatory elements that control the FIX gene expression in these cassettes were screened for liver-specific effectiveness. Among all the constructed cassettes, scAAV-Pre-hFIXco-SIH-R338L, which was the construct under the control of the prothrombin enhancer and prealbumin promoter, resulted in the highest level of coagulant activity, and the expression levels of two constructed cassettes （scAAV-Chi-hFIXco-SIH-R338L and scAAV-Pre- hFIXco-SIH-R338L） were also higher than that of the published cassette （scAAV-LPI-hFIXco-SJ）. In summary, our strategies led to a substantial increase in hFIX expression at the protein level or a remarkably elevated coagulant activity. Thus, these reconstructs of hFIX with AAV vector may potentially contribute to the creation of an efficacious gene therapy of hemophilia B.