Bottom-up assembly of RNA  nanoparticles containing phi29 motor pRNA to silence the asthma STAT5b gene.
Activation of the transcription factor signal transducer and activator of transcription 5b (STAT5b) is a key event in the development of asthma. The potent ability of small interfering RNA (siRNA) to inhibit the expression of STAT5b mRNA has provided a new class of therapeutics for asthma. However, efficient delivery of siRNAs remains a key obstacle to their successful application. A targeted intracellular delivery approach for siRNA to specific cell types would be highly desirable. We used packaging RNA (pRNA), a component of the bacteriophage phi29-packaging motor, to deliver STAT5b siRNA to asthmatic spleen lymphocytes. This pRNA was able to spontaneously carry siRNA/STAT5b and aptamer/CD4, which is a ligand to CD4 molecule. Based on RT-PCR data, the pRNA dimer effectively inhibited STAT5b gene mRNA expression of asthmatic spleen lymphocytes, without the need for additional transfections. We conclude that the pRNA dimer carrying both siRNA and aptamer can deliver functional siRNA to cells; possibly, the aptamer acts as a ligand to interact with specific receptors. The pRNAs were evaluated with a CCK-8 kit and were found to have little cytotoxicity. We conclude that pRNA as a novel nanovehicle for RNA worth further study.
Department of Respiratory Diseases, Second Clinical Medical College, Jinan University, Shenzhen, Guangdong, P.R. China. dalyschloe@yahoo.cn
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D000818:Animals; D001249:Asthma / Q000235:genetics* / Q000473:pathology; D017103:Bacillus Phages / Q000235:genetics*; D001483:Base Sequence; D016923:Cell Death; D019281:Dimerization; D005260:Female; D020868:Gene Silencing; D008168:Lung / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D034741:RNA, Small Interfering / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050799:STAT5 Transcription Factor / Q000235:genetics*; D012844:Sincalide; D019065:Virus Assembly / Q000235:genetics*
D034741:RNA, Small Interfering; D012367:RNA, Viral; D050799:STAT5 Transcription Factor; D012844:Sincalide