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• W3025682495 abstract "To explore the feasibility of co-transduction and co-expression of Nogo extracellular peptide residues 1-40 (NEP1-40) gene and neurotrophin 3 (NT-3) gene into neural stem cells (NSCs).NSCs were derived from the cortex tissue of Sprague Dawley rat embryo. The experiment included 5 groups: no-load lentiviral vector transducted NSCs (group A), NEP1-40 transducted NSCs (group B), NT-3 transducted NSCs (group C), NEP1-40 and NT-3 corporately transducted NSCs (group D), and blank control (group E). Target genes were transducted into NSCs by lentiviral vectors of different multiplicity of infection (MOI; 5, 10, 15) for different time (24, 48, 72 hours). Fluorescent microscope was used to observe the expression of fluorescence protein and acquire the optimum MOI and optimum collection time. Real-time fluorescence quantitative PCR and Western blot tests were utilized to evaluate the gene expressions of NEP1-40 and NT-3 in NSCs and protein expressions of NEP1-40 and NT-3 in NSCs and in culture medium.The optimum MOI for both target gene was 10 and the optimum collection time was 48 hours. The real-time fluorescence quantitative PCR and Western blot results showed that the mRNA and protein relative expressions of NEP1-40 in groups B and D were significantly higher than those in groups A and C ( P<0.05), but no significant difference was found between groups B and D, and between groups A and C ( P>0.05). The mRNA and protein relative expressions of NT-3 in groups C and D were significantly higher than those in groups A and B ( P<0.05), but no significant difference was found between groups A and B, and between groups C and D ( P>0.05).NEP1-40 and NT-3 gene can be successfully co-transducted into NSCs by the mediation of lentiviral vector. The expressions of the two target genes are stable and have no auxo-action or antagonism between each other.通过慢病毒载体将 NEP1-40（Nogo extracellular peptide residues 1-40）及神经营养因子 3（neurotrophin 3，NT-3）双基因转染入神经干细胞（neural stem cells，NSCs）内进行表达，探讨 NEP1-40 及 NT-3 双基因转染 NSCs 的可行性，为 NSCs 体内实验奠定基础。.将 SD 大鼠胚胎室管膜区 NSCs 采用空载慢病毒载体（A 组）、NEP1-40 慢病毒载体（B 组）、NT-3 慢病毒载体（C 组）及 NEP1-40 和 NT-3 慢病毒载体（D 组）进行转染，以未转染病毒的细胞作为对照组（E 组）。用感染复数（multiplicity of infection，MOI）为 5、10、15 的慢病毒载体分别转染 24、48、72 h，荧光显微镜观察转染后细胞内荧光表达情况，确定慢病毒载体的最佳 MOI 和收样时间。再分别通过实时荧光定量 PCR 及 Western blot，检测转染后细胞中 NEP1-40 及 NT-3 基因的表达，以及细胞和培养基中 NEP1-40 及 NT-3 蛋白的表达。.荧光显微镜观察示，MOI 为 10 时 NEP1-40 和 NT-3 基因慢病毒载体在 NSCs 内转染率最高，最佳时间为转染 48 h 时。实时荧光定量 PCR 及 Western blot 检测示，B、D 组 NEP1-40 mRNA 相对表达量和蛋白相对表达量均显著高于 A、C 组，差异有统计学意义（ P<0.05）；A、C 组间及 B、D 组间比较差异无统计学意义（ P>0.05）。C、D 组 NT-3 mRNA 相对表达量和蛋白相对表达量均显著高于 A、B 组，差异有统计学意义（ P<0.05）；A、B 组间和 C、D 组间比较差异无统计学意义（ P>0.05）。.通过慢病毒载体可将 NEP1-40 及 NT-3 双基因成功转染入 NSCs 内，在 NSCs 内稳定表达，且两种目的基因在表达过程中无相互拮抗或促进作用。." @default.
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• W3025682495 date "2018-04-15" @default.
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• W3025682495 title "[Experimental study of lentivirus-mediated Nogo extracellular peptide residues 1-40 gene and neurotrophin 3 gene co-transduction in neural stem cells]." @default.
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• W3025682495 doi "https://doi.org/10.7507/1002-1892.201710079" @default.
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