Blagbrough, I. S., Metwally, A. A. and Ghonaim, H. M. (2012) Asymmetrical N4,N9-Diacyl Spermines: SAR Studies of Nonviral Lipopolyamine Vectors for Efficient siRNA Delivery with Silencing of EGFP
نویسندگان
چکیده
Our aim is to study the effects of varying the two acyl moieties in synthesized N,N-diacyl spermines on siRNA formulations and their delivery efficiency in cell lines. Six novel asymmetrical lipopolyamines: [N-cholesteryloxy-3-carbonyl-N-oleoyl-, N-decanoyl-N-oleoyl-, N-decanoyl-N-stearoyl-, N-lithocholoyl-N-oleoyl-, N-myristoleoyl-N-myristoyl-, and N oleoyl-N-stearoyl]-1,12-diamino-4,9-diazadodecane were assessed for their abilities to bind to siRNA, studied using a RiboGreen intercalation assay, and to form nanoparticles. Their siRNA delivery efficiencies were quantified in FEK4 primary skin cells and in an immortalized cancer cell line (HtTA) using a fluorescein-tagged siRNA, and compared with formulations of N,N dioleoyl-1,12-diamino-4,9-diazadodecane and of a leading transfecting agent, TransIT-TKO. Transfection was measured in terms of siRNA delivery and silencing of EGFP reporter gene in HeLa cells. By incorporating two different acyl moieties, changing their length and oxidation level in a controlled manner, we show efficient fluorescein-tagged siRNA formulation, delivery, and knock-down of EGFP reporter gene. N-Oleoyl-N-stearoyl spermine and N-myristoleoyl-N myristoyl spermine are effective siRNA delivery vectors typically resulting in 89% cell delivery and gene silencing to 34% in the presence of serum, comparable with the results obtained with TransIT-TKO; adding a second lipid chain is better than incorporating a steroid moiety.
منابع مشابه
Efficient silencing of EGFP reporter gene with siRNA delivered by asymmetrical N4,N9-diacyl spermines.
It is important to obtain structure-activity relationship (SAR) data across cationic lipids for the self-assembly and nonviral intracellular delivery of siRNA. The aims of this work are to carry out a SAR study on the efficiency of asymmetrical N(4),N(9)-diacyl spermines in siRNA delivery and EGFP reporter gene silencing, with comparisons to selected mixtures composed of symmetrical N(4),N(9)-d...
متن کاملMolecular Pharmaceutics, 2012, 9, 1853-1861 Asymmetrical N,N-Diacyl Spermines: SAR Studies of Non-Viral Lipopolyamine Vectors for Efficient siRNA Delivery with Silencing of EGFP Reporter Gene
Our aim is to study the effects of varying the two acyl moieties in synthesized N,N-diacyl spermines on siRNA formulations and their delivery efficiency in cell lines. Six novel asymmetrical lipopolyamines: [N-cholesteryloxy-3-carbonyl-N-oleoyl-, N-decanoyl-N-oleoyl-, N-decanoyl-N-stearoyl-, N-lithocholoyl-N-oleoyl-, N-myristoleoyl-N-myristoyl-, and N oleoyl-N-stearoyl]-1,12-diamino-4,9-diazad...
متن کاملMolecular Pharmaceutics, 2012, 9, 1862-1876 Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N,N-Diacyl Spermines
It is important to obtain structure-activity relationship (SAR) data across cationic lipids for the self-assembly and non-viral intracellular delivery of siRNA. The aims of this work are to carry out a SAR study on the efficiency of asymmetrical N,N-diacyl spermines in siRNA delivery and EGFP reporter gene silencing, with comparisons to selected mixtures composed of symmetrical N,N-diacyl sperm...
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Four guanidine derivatives of N4,N9-diacylated spermine have been designed, synthesized, and characterized. These guanidine-containing cationic lipids bound siRNA and formed nanoparticles. Two cationic lipids with C18 unsaturated chains, N1,N12-diamidino-N4,N9-dioleoylspermine and N1,N12-diamidino-N4-linoleoyl-N9-oleoylspermine, were more efficient in terms of GFP expression reduction compared ...
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Gene silencing by siRNA (synthetic dsRNA of 21-25 nucleotides) is a well established biological tool in gene expression studies and has a promising therapeutic potential for difficult-to-treat diseases. Five fatty acids of various chain length and oxidation state (C12:0, C18:0, C18:1, C18:2, C22:1) were conjugated to the naturally occurring polyamine, spermine, and evaluated for siRNA delivery ...
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