Construction of microRNA expression report vector targeting CD4 and functional research of the vector / 中华微生物学和免疫学杂志

ABSTRACT

ObjectiveTo construct expressing vector of microRNA with molecular cloning methods which target CD4 and electroporating the vector to the MT4 cell to determine its effect to CD4 expression.MethodsPredict a microRNA can target CD4.Linking the pre-mir-181a PCR products to T vector,then cloning into the pEGFP-N1 vector after enzyme digestion.To test the integrity through the colony PCR and sequencing analysis.Electroporating the vector to MT4 cell,using FACS to test the CD4 expression.ResultsHsa-mir-181a is able to target CD4.The sequence of the construct was correct.The hsa-mir-181a is stable expressing in MT4 after electroporating with the vector and MT4 cell CD4 was down-regulated.ConclusionThe construct can be stable expressing hsa-mir-181a in MT4 cell and down-regulating the CD4 expressing.This method can be utilized as a novel intervention to the HIV fusion,it shows potential as a gene therapy tool in vitro.