【摘要】 应用体外转录和体外剪接系统，以体外转录产生的特异性反义RNA封闭IVS－2－654→T突变基因中3’隐匿剪接位点和5’异常剪接位点，使之失去活性，以减少异常加工的mRNA，且部分恢复了正常剪接途径，其正常剪接mRNA的水平［β／（β十β）］从处理前的0．25上升到处理后的0．46—0．61。研究结果提示了应用反义核酸临床治疗β－地贫（IVS－2－654C→T突变）的可能性。更多还原

【Abstract】 The C→T substitution at position IVS-2-nt. 654 in the β-globin gene, a mutation causing defect in RNA splicing, comprises one of the most common β-thalassaemia alleies in Chinese β-thalassaemia patients.In this study, three suitable IN-Vitro-transcribed antisense RNAs of Al, A2 and A3 have been designed to target against the cryptic 3’ splice site at position IVS-2-nt. 579 and the aberrant 5’ splice site at position IVS-2-nt. 654 respectively, and allow them to reverse incorrect splicing of β-thalassaemic pre-mRNA. The results show that the ratio of correct splicing mRNA(β) to total mRNA (β+β ) is increased from 0. 25 to 0. 46, 0. 60, 0. 61 with treatment of Al, A2, A3, indicating that the above three antisense RNAs can restore correct splicing in thalassaemic β-globin (IVS-2-654 C→T) pre-mRNA by blocking aberrant splicing sites, and forcing the splicing mechanism to reselect correct splicing sites. Therefore, the technology of antisense RNA provides an additional approach to the climical therapy of β-thalassaemia.更多还原