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Development of a targeted siRNA delivery system using FOL-PEG-PEI conjugate
Published in
2010
PMID: 19816791
Volume: 37
   
Issue: 6
Pages: 2919 - 2926
Abstract
Receptor mediated delivery of siRNA enables silencing of target genes in specific tissues. Folate receptor (FR) is an attractive target for tumor-selective gene delivery. The focus of this study was to deliver the dihydrofolate reductase (DHFR) siRNA expressing plasmid and to silence the DHFR gene in FR positive KB cells, by complexing the plasmid with a folate-polyethylene glycol-polyethylenimine (FOL-PEG-PEI) conjugate, as a gene carrier. A DHFR siRNA sequence was cloned into a pSUPER-RNAi vector and complexed with the FOL-PEG-PEI conjugate. The complex was characterized by particle size analyzer, gel retardation and DNase protection assay. The FOL-PEG-PEI/pSUPER-siDHFR complex was transfected to FR overexpressing (KB) and FR negative (A549) cells. The transfection effiencies and gene inhibition were analyzed by fluorescence microscopy and RT-PCR. The pSUPER-siDHFR/PEI-PEG-FOL complex delivered the siRNA vector and inhibited DHFR gene in KB cells, while A549 cells were unaffected. Lipofectamine mediated transfection of pSUPER-siDHFR, delivered the vector and inhibited the DHFR gene in both KB and A549 cells. FR mediated delivery of siDHFR complexed with PEI-PEG-FOL conjugate inhibits the DHFR expression in FR positive cells alone. This strategy can be extended to deliver a wide range of drugs and post-transcriptional gene silencing therapeutics. © 2009 Springer Science+Business Media B.V.
About the journal
JournalMolecular Biology Reports
ISSN03014851
Open AccessNo
Concepts (54)
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    Dihydrofolate reductase
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    Folate receptor
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    Folic acid
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    LIPOFECTAMINE
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    Macrogol
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    Polyethyleneimine
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    Small interfering rna
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    DEOXYRIBONUCLEASE I
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    Macrogol derivative
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    Analyzer
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    Article
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    CELL STRAIN KB
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    Controlled study
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    Drug delivery system
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    Enzyme assay
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    Fluorescence microscopy
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    Gene delivery system
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    Gene targeting
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    GENE VECTOR
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    Genetic transfection
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    Human
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    Human cell
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    Molecular cloning
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    Plasmid
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    POSTTRANSCRIPTIONAL GENE SILENCING
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    Reverse transcription polymerase chain reaction
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    Rna interference
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    RNA SEQUENCE
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    Gene transfer
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    Genetics
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    Metabolism
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    Molecular genetics
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    NUCLEASE PROTECTION ASSAY
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    Nucleotide sequence
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    Tumor cell line
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    Base sequence
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    Cell line, tumor
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    Cloning, molecular
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    DEOXYRIBONUCLEASE I
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    Gene transfer techniques
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    GENETIC VECTORS
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    Humans
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    Microscopy, fluorescence
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    Molecular sequence data
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    NUCLEASE PROTECTION ASSAYS
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    Polyethylene glycols
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    Reverse transcriptase polymerase chain reaction
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    Rna, small interfering
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    TETRAHYDROFOLATE DEHYDROGENASE
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    Transfection
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    DEOXYRIBONUCLEASE I
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    GENETIC VECTORS
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    NUCLEASE PROTECTION ASSAYS
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    TETRAHYDROFOLATE DEHYDROGENASE