摘要
目的:筛选有效抑制大鼠CRMP1基因表达的siRNA序列。方法:化学合成3对针对大鼠CRMP1基因的特异性siRNA,用脂质转染胺(lipofectamine)LTX转染大鼠海马神经元,通过流式细胞仪检测转染效率,使用RT-PCR与实时PCR检测CRMP1基因mRNA的表达,从而筛选抑制效率最高的siRNA。结果:流式细胞仪检测转染效率为52.5%。通过RT-PCR的初步检测和实时PCR的定量检测结果显示,与对照组相比,所设计的3对siRNA均能不同程度抑制CRMP1基因mRNA的表达,其中CRMP1-249 siRNA抑制效果最佳,抑制率达84.3%。结论:成功筛选出1对能有效抑制大鼠CRMP1基因表达的siRNA序列。
Aim：To screen small interfering RNA（siRNA） sequences with CRMP1 mRNA in rat hippocampal neurons. Methods： After siRNAs for CRMP1 gene were designed and synthesized,they were transfected into rat hippocampal neurons by positive ion liposome lipofectamine LTX.Transfected efficiency was assessed by flow cytometry,and RT-PCR and real-time PCR were used to detect the expression of CRMP1 mRNA.Results： The transfected efficiency was 52.5% detected by flow cytometry.From the results of RT-PCR and real-time PCR,we found that all of three couples of siRNAs customized to silence target-gene,which could inhibit gene expression of CRMP1,compared with the control group.CRMP1-249,one of three couples of siRNAs,had the best inhibitory effect on gene expression of CRMP1,and the inhibitory efficiency was 84.3%.Conclusion：The present experiment had successfully screened a couple of synthetic siRNA Sequence to CRMP1 mRNA.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2010年第6期560-564,共5页
Journal of Jinan University(Natural Science & Medicine Edition)
基金
广东省自然科学基金项目(8451063201000193)
广东省科技计划项目(2010B031600102)
广东省医学科研基金项目(A2008344)
