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目的究设计一种病毒载体介导的体内调节系统,依据诱导性Cre重组酶来调控转基因治疗帕金森病大鼠的基因表达。方法将重组腺相关病毒(AAV)载体表达的Cre重组酶融合到雌激素受体的配体结合区,与AAV载体表达的多巴胺合成酶一起转导到帕金森病大鼠模型中。用合成的雌激素受体调节剂他莫昔芬处理。结果诱导位于LoxP侧面的酪氨酸羟化酶(TH)序列选择性的敲掉,导致多巴胺的合成减少,而芳香族氨基酸脱羧酶(AADC)的表达不受影响,从而保持左旋多巴的疗效。结论病毒载体介导的诱导性的Cre重组酶在体内的应用可作为一种分子开关,对转基因表达进行时空调控,从而增加基因治疗的安全性。
OBJECTIVE: To design a viral vector-mediated in vivo regulatory system to regulate the gene expression in transgenic Parkinson’s disease rats by induction of Cre recombinase. Methods Recombinant adeno-associated virus (AAV) vector expressing Cre recombinase was fused to the ligand-binding region of estrogen receptor and transduced into Parkinson’s disease rat model with dopamine synthase expressed by AAV vector. Treatment with the synthetic estrogen receptor modulator tamoxifen. As a result, tyrosine hydroxylase (TH) sequence on the side of LoxP was selectively knocked out, resulting in a decrease of dopamine synthesis, while the expression of AADC was not affected, thereby maintaining the efficacy of levodopa . Conclusion The use of viral vector-mediated inducible Cre recombinase in vivo can act as a molecular switch to spatiotemporal regulation of transgene expression, thereby increasing the safety of gene therapy.