Development of a microRNA delivery system based on bacteriophage MS2 virus‐like particles

Recently, microRNA (miRNA)‐mediated RNA interference has been developed as a useful tool in gene function analysis and gene therapy. A major obstacle in miRNA‐mediated RNAi is cellular delivery, which requires an efficient and flexible delivery system. The self‐assembly of the MS2 bacteriophage capsids has been used to develop virus‐like particles (VLPs) for RNA and drug delivery. However, MS2 VLP‐mediated miRNA delivery has not yet been reported. We therefore used an Escherichia coli expression system to produce the pre‐miR 146a contained MS2 VLPs, and then conjugated these particles with HIV‐1 Tat47–57 peptide. The conjugated MS2 VLPs effectively transferred the packaged pre‐miR146a RNA into various cells and tissues, with 0.92–14.76‐fold higher expression of miR‐146a in vitro and about two‐fold higher expression in vivo, and subsequently suppressed its targeting gene. These findings suggest that MS2 VLPs can be used as a novel vehicle in miRNA delivery systems, and may have applications in gene therapy.

[1]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[2]  T. Rana,et al.  Visualizing a correlation between siRNA localization, cellular uptake, and RNAi in living cells. , 2004, Chemistry & biology.

[3]  Anil K Sood,et al.  Therapeutic EphA2 gene targeting in vivo using neutral liposomal small interfering RNA delivery. , 2005, Cancer research.

[4]  Eric J Wagner,et al.  Both natural and designed micro RNAs can inhibit the expression of cognate mRNAs when expressed in human cells. , 2002, Molecular cell.

[5]  Phillip D. Zamore,et al.  RNA Interference , 2000, Science.

[6]  Beverly L. Davidson,et al.  Current prospects for RNA interference-based therapies , 2011, Nature Reviews Genetics.

[7]  A. Bader,et al.  Developing therapeutic microRNAs for cancer , 2011, Gene Therapy.

[8]  Jinming Li,et al.  A new RNA vaccine platform based on MS2 virus-like particles produced in Saccharomyces cerevisiae. , 2011, Biochemical and biophysical research communications.

[9]  S. Lowe,et al.  A microRNA polycistron as a potential human oncogene , 2005, Nature.

[10]  K. Livak,et al.  Real-time quantification of microRNAs by stem–loop RT–PCR , 2005, Nucleic acids research.

[11]  Chris J. Adams,et al.  RNA Bacteriophage Capsid-Mediated Drug Delivery and Epitope Presentation , 2003, Intervirology.

[12]  M. Sakagami,et al.  In vivo, in vitro and ex vivo models to assess pulmonary absorption and disposition of inhaled therapeutics for systemic delivery. , 2006, Advanced drug delivery reviews.

[13]  J Barsoum,et al.  Tat-mediated delivery of heterologous proteins into cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[14]  V. Ambros,et al.  An Extensive Class of Small RNAs in Caenorhabditis elegans , 2001, Science.

[15]  J M Walker,et al.  The bicinchoninic acid (BCA) assay for protein quantitation. , 1994, Methods in molecular biology.

[16]  M. Carmell,et al.  Posttranscriptional Gene Silencing in Plants , 2006 .

[17]  G. Hannon.  RNA interference : RNA , 2002 .

[18]  P. Tak,et al.  MicroRNA-146A contributes to abnormal activation of the type I interferon pathway in human lupus by targeting the key signaling proteins. , 2009, Arthritis and rheumatism.

[19]  Xi-sha Chen,et al.  MicroRNA‐146a and Human Disease , 2010, Scandinavian journal of immunology.

[20]  Stephen A. Bustin,et al.  Real-Time PCR , 2005 .

[21]  W. Brown,et al.  Delivery of antisense oligonucleotides to leukemia cells by RNA bacteriophage capsids. , 2005, Nanomedicine : nanotechnology, biology, and medicine.

[22]  Mark B. Carter,et al.  Cell-specific delivery of diverse cargos by bacteriophage MS2 virus-like particles. , 2011, ACS nano.

[23]  R. Ketting.  microRNA Biogenesis and Function : An overview. , 2011, Advances in experimental medicine and biology.

[24]  Paul J. Hertzog,et al.  Analysis of microRNA turnover in mammalian cells following Dicer1 ablation , 2011, Nucleic acids research.

[25]  D. Maclean,et al.  Therapeutic applications of cell-penetrating peptides. , 2011, Methods in molecular biology.

[26]  D. Bartsch,et al.  Quantitative analysis of conditional gene inactivation using rationally designed, tetracycline-controlled miRNAs , 2010, Nucleic acids research.

[27]  S. Kauppinen,et al.  LNA-modified oligonucleotides mediate specific inhibition of microRNA function. , 2006, Gene.

[28]  J. Au,et al.  Delivery of siRNA Therapeutics: Barriers and Carriers , 2010, The AAPS Journal.

[29]  Carl O. Pabo,et al.  Cellular uptake of the tat protein from human immunodeficiency virus , 1988, Cell.

[30]  K. Channon,et al.  RNAi-mediated knockdown of HMG CoA reductase enhances gene expression from physiologically regulated low-density lipoprotein receptor therapeutic vectors in vivo , 2011, Gene Therapy.

[31]  A. Pasquinelli,et al.  Regulation by let-7 and lin-4 miRNAs Results in Target mRNA Degradation , 2005, Cell.

[32]  Kuo Zhang,et al.  Armored Long RNA Controls or Standards for Branched DNA Assay for Detection of Human Immunodeficiency Virus Type 1 , 2009, Journal of Clinical Microbiology.

[33]  P. Stockley,et al.  Molecular mechanism of RNA-phage morphogenesis. , 1993, Biochemical Society transactions.

[34]  Jerry Pelletier,et al.  Short RNAs repress translation after initiation in mammalian cells. , 2006, Molecular cell.

[35]  Sung-Hoon Kim,et al.  Cytoplasmic transduction peptide (CTP): new approach for the delivery of biomolecules into cytoplasm in vitro and in vivo. , 2006, Experimental cell research.

[36]  Thomas D. Schmittgen,et al.  Real-time PCR quantification of precursor and mature microRNA. , 2008, Methods.

[37]  P. Linsley,et al.  miR-146a is a significant brake on autoimmunity, myeloproliferation, and cancer in mice , 2011, The Journal of experimental medicine.

[38]  Wei Wang,et al.  Development of an antisense RNA delivery system using conjugates of the MS2 bacteriophage capsids and HIV-1 TAT cell-penetrating peptide. , 2009, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[39]  Kaleb M. Pauley,et al.  miR-146a Is Critical for Endotoxin-induced Tolerance , 2009, The Journal of Biological Chemistry.