The identification of human microRNA335′s predicated target gene CCL1 1,CCL26 and SOX4
10.3969/j.issn.1671-8348.2015.24.002
- VernacularTitle:人 MicroRNA335的预测靶基因 CCL11 CCL26及 SOX4的鉴定
- Author:
Zhihong WEN
;
Yan DAI
;
Shuang HE
- Publication Type:Journal Article
- Keywords:
MicroRNAs;
hsa-miR-335;
target gene;
gene regulation;
CCL1 1;
CCL26;
SOX4;
dual luciferase reporter gene sys-tem
- From:
Chongqing Medicine
2015;(24):3316-3318
- CountryChina
- Language:Chinese
-
Abstract:
Objective To identify the targeted-regulating relationship between human MicroRNA335 (hsa-miR-335 )and CCL1 1,CCL26 and SOX4.Methods The potential fragments of hsa-miR-335 target genes CCL1 1,CCL26 and SOX4 were predicted by the bioinformatics analyzing tools online.The 3′untranslated regions(3′UTR)of the CCL1 1,CCL26 and SOX4 were connected to the eukaryotic expression vectors pMIR REPORT.The constructs of pMIR-REPORT-CCL1 13′UTR,pMIR-REPORT-CCL26 3′UTR,pMIR-REPORT-SOX4 3′UTR and positive control were co-transfected with Pre-miRTM miRNA335 Precursor or negative control into 293 T7/1 7 cell line by lipofectamine 2000,respectively.Both Firefly and Renilla luciferase activity were detected by dual luciferase reporter assay system.Results Compared with the negative control group,luciferase assay revealed that has-miR-335 could significantly diminish luciferase activity from SOX4 reporter vector (P <0.01 ),while the suppression of luciferase activity was not found in CCL1 1 or CCL26 reporter vector (P >0.05).Conclusion The results suggested that hsa-miR-335 targeted regu-lated SOX4,but not targeted CCL1 1 and CCL26.
