PMID: 30014628Jul 18, 2018Paper

Investigation of Lentiviral Vectors Based Integrin β8 RNAi System in Neonatal Rats' Brain

Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition
Bin-Zhi TangDe-Jian Wang

Abstract

To construct lentiviral vectors expressing pSicoR-β8 shRNA and evaluate its efficiency of RNA interference in neonatal rats' brain. Plasmid vectors pSicoR-β8 shRNA and pSicoR-control,as well as lentiviral packaging system pDM2G,g/p RRE and pRSV Rev were amplified respectively and plasmid DNA was identified by restriction enzyme digestion. Lentiviral packaging system and expressing vector pSicoR-β8 shRNA/pSicoR-control were co-transfected into packaging cell line 293T. Lentiviral particles expressing β8-shRNA or control sequence packaged and secreted by 293T were collected,concentrated by PEG-it,and viral titers were assayed by 50% tissue culture infective dose (TCID50). RNAi for integrin β8 in neonatal rats' brain was performed by intraventricular injection of lentivirus expressing β8-shRNA and rats received lentivirus expressing β8-shRNA were served as control. Green fluorescent protein (GFP) expression after intraventricular injection of GFP-Lentivirus was observed under fluorescence microscope,β8 mRNA and β8 protein expression were detected by RT-PCR and Western blot respectively,all of which were performed to evaluate the RNAi efficiency and to choose the optimal time for intervention. Restrictive endonuclease digestion and...Continue Reading

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