Methods in molecular biology (Clifton, N.J.), 2011, Vol.683, p.339
2011
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- Autor(en) / Beteiligte
- Titel
- PTD-DRBD siRNA delivery
- Ist Teil von
- Methods in molecular biology (Clifton, N.J.), 2011, Vol.683, p.339
- Ort / Verlag
- United States
- Erscheinungsjahr
- 2011
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- A major hurdle in drug delivery today is for the drug to reach inside the cell to exert its biological effect. Many drug candidates are hydrophilic and are therefore not able to cross the hydrophobic plasma membrane, which serves to protect the cell from foreign molecules and pathogens. One promising drug candidate is the hydrophilic and negatively charged short-interfering RNA (siRNA), known to degrade target mRNA 1,000-fold more efficiently than small molecule drugs. The delivery capacity of small cationic peptides called protein transduction domains or cell-penetrating peptides, suggested them to be suitable delivery vehicles for siRNA. However, it has proven troublesome to utilize the PTD-siRNA conjugates for mRNA degradation due to the characteristics of siRNA, often resulting in precipitation and aggregation. This chapter describes a recently reported delivery strategy, PTD-DRBD fusion protein siRNA delivery, where a double-stranded RNA-binding domain expressed as a fusion protein together with three TAT PTDs binds the siRNA, thus masking the negatively charged backbone and preventing aggregation. This new protocol results in noncytotoxic mRNA degradation even more effective than lipofection.
- Sprache
- Englisch
- Identifikatoren
- eISSN: 1940-6029DOI: 10.1007/978-1-60761-919-2_24Titel-ID: cdi_pubmed_primary_21053141
- Format
- –
- Schlagworte
- Cell Line, Cell-Penetrating Peptides - genetics, Cell-Penetrating Peptides - isolation & purification, Cell-Penetrating Peptides - metabolism, Drug Carriers - isolation & purification, Drug Carriers - metabolism, Flow Cytometry, Gene Knockdown Techniques, Humans, Immunohistochemistry, Interferon-alpha - metabolism, Oligonucleotide Array Sequence Analysis, Protein Engineering, Protein Structure, Tertiary, Recombinant Fusion Proteins - genetics, Recombinant Fusion Proteins - isolation & purification, Recombinant Fusion Proteins - metabolism, Reverse Transcriptase Polymerase Chain Reaction, RNA, Double-Stranded - metabolism, RNA, Small Interfering - genetics, RNA, Small Interfering - metabolism, Transfection - methods, Tumor Necrosis Factor-alpha - metabolism