GAS41基因RNA干扰慢病毒载体的构建

Construction of shRNA lentiviral vector for targeting GAS41 gene

目的 :构建GAS41-shRNA慢病毒载体。方法 :合成GAS41干扰序列,构建GV118-GAS41-RNAi慢病毒干扰载体,连同包装质粒p Helper1.0和包膜质粒p Helper2.0共同转染293T细胞,收集储存病毒颗粒并进行滴度测定。结果 :构建的GAS41干扰慢病毒载体GV118-GAS41-RNAi经PCR和测序鉴定,结果与设计序列相符,收获病毒颗粒,测定滴度为5×108TU/m L。结论 :成功构建了GAS41-shRNA慢病毒载体,为后续进一步研究GAS41基因在视神经胶质瘤(optic nerve glioma,ONG)中的作用奠定了基础。

Objective To build glioma amplified sequence 41(GAS41) interference lentiviral vectors. Methods The GV118-GAS41-RNAi vectors were cotransfected into 293 T incasing cells with p Helper 1.0 and p Helper 2.0. Lentiviruses were harvested and viral titer was detected. Results The recombinant lentiviral vectors were successfully constructed and identified by PCR and DNA sequencing. The lentiviruses were harvested from supernatant of 293 T cells with a viral titer of 5×108 TU/m L. Conclusion The interference lentivirus vectors of GAS41 have been successfully constructed, which will lay the foundation for the following study of GAS41 gene in optic nerve glioma(ONG).