趋化因子受体基因CCR5人工miRNA的构建与功能分析

发布时间：2018-02-08 17:28

本文关键词： HIV- CCR基因 RNA干扰人工miRNA　出处：《中国皮肤性病学杂志》2017年04期 　论文类型：期刊论文

【摘要】：目的构建人工miR-CCR5,并研究其对HIV-1感染宿主细胞的影响。方法以miR-155为基础骨架,根据HIV-1 CCR5基因序列设计并合成4对miRNA寡聚单链DNA,构建4个人工miR-CCR5。采用qPCR技术检测其对CCR5基因的沉默效率和对干扰素表达的影响;MTT法测定其对被转染细胞的毒性;抗HIV-1_(BaL)感染实验检测其对HIV-1感染的抑制作用。结果 qPCR结果显示,4个人工miR-CCR5对CCR5基因都具有一定的沉默效果,其中miR-CCR5-4的沉默效率最高,达72%;且不会对细胞干扰素的表达产生影响。MTT实验证实miR-CCR5-4不会影响被转染细胞的活性,此外,病毒抑制实验显示,miR-CCR5-4对HIV-1_(BaL)p24蛋白的抑制作用达70.48%,效果明显。结论所获得的miR-CCR5-4可为进一步的抗HIV-1_(BaL)感染研究提供基础。
[Abstract]:Objective to construct artificial miR-CCR5 and study the effect of miR-CCR5 on HIV-1 infected host cells. According to the sequence of HIV-1 CCR5 gene, four pairs of miRNA oligonucleotides were designed and synthesized, and four artificial miR-CCR5s were constructed. The silencing efficiency of CCR5 gene and the expression of interferon were detected by qPCR technique. Results the results of qPCR showed that the four artificial miR-CCR5 had certain silencing effect on CCR5 gene, and miR-CCR5-4 had the highest silencing efficiency. No effect on the expression of interferon. MTT assay confirmed that miR-CCR5-4 did not affect the activity of transfected cells. Virus inhibition test showed that the inhibitory effect of miR-CCR5-4 on HIV-1_(BaL)p24 protein was 70.48. Conclusion the obtained miR-CCR5-4 may provide a basis for further research on anti-HIV-1 Bal infection.
【作者单位】：衡阳师范学院生命科学与环境学院功能金属有机材料湖南省重点实验室;南华大学药学与生命科学学院;Laboratory
【基金】：湖南省自然科学基金重点项目(09JJ3034)
【分类号】：R512.91

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