title: Cationic amphiphilic macromolecule (CAM)-lipid complexes for efficient siRNA gene silencing.
abstract: The accumulated evidence has shown that lipids and polymers each have distinct advantages as carriers for siRNA delivery. Composite materials comprising both lipids and polymers may present improved properties that combine the advantage of each. Cationic amphiphilic macromolecules (CAMs) containing a hydrophobic alkylated mucic acid segment and a hydrophilic poly(ethylene glycol) (PEG) tail were non-covalently complexed with two lipids, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), to serve as a siRNA delivery vehicle. By varying the weight ratio of CAM to lipid, cationic complexes with varying compositions were obtained in aqueous media and their properties evaluated. CAM-lipid complex sizes were relatively independent of composition, ranging from 100 to 200nm, and zeta potentials varied from 10 to 30mV. Transmission electron microscopy confirmed the spherical morphology of the complexes. The optimal N/P ratio was 50 as determined by electrophoretic mobility shift assay. The ability to achieve gene silencing was evaluated by anti-luciferase siRNA delivery to a U87-luciferase cell line. Several weight ratios of CAM-lipid complexes were found to have similar delivery efficiency compared to the gold standard, Lipofectamine. Isothermal titration calorimetry revealed that siRNA binds more tightly at pH=7.4 than pH=5 to CAM-lipid (1:10 w/w). Further intracellular trafficking studies monitored the siRNA escape from the endosomes at 24h following transfection of cells. The findings in the paper indicate that CAM-lipid complexes can serve as a novel and efficient siRNA delivery vehicle.
authors.affiliation: Rutgers, The State University of New Jersey, Department of Chemistry and Chemical Biology, 610 Taylor Road, Piscataway, NJ 08854, United States.
authors.forename: Li
authors.initials: L
authors.lastname: Gu
authors.affiliation: Rutgers, The State University of New Jersey, Department of Biomedical Engineering, 599 Taylor Road, Piscataway, NJ 08854, United States.
authors.forename: Leora M
authors.initials: LM
authors.lastname: Nusblat
authors.affiliation: Yeshiva University, Stern College for Women, Department of Chemistry and Biochemistry, New York, NY 10016, United States.
authors.forename: Nasim
authors.initials: N
authors.lastname: Tishbi
authors.affiliation: Yeshiva University, Stern College for Women, Department of Chemistry and Biochemistry, New York, NY 10016, United States.
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Noble
authors.affiliation: Yeshiva University, Stern College for Women, Department of Chemistry and Biochemistry, New York, NY 10016, United States.
authors.forename: Chaya M
authors.initials: CM
authors.lastname: Pinson
authors.affiliation: Yeshiva University, Stern College for Women, Department of Chemistry and Biochemistry, New York, NY 10016, United States.
authors.forename: Evan
authors.initials: E
authors.lastname: Mintzer
authors.affiliation: Rutgers, The State University of New Jersey, Department of Biomedical Engineering, 599 Taylor Road, Piscataway, NJ 08854, United States.
authors.forename: Charles M
authors.initials: CM
authors.lastname: Roth
authors.affiliation: Rutgers, The State University of New Jersey, Department of Chemistry and Chemical Biology, 610 Taylor Road, Piscataway, NJ 08854, United States; Rutgers, The State University of New Jersey, Department of Biomedical Engineering, 599 Taylor Road, Piscataway, NJ 08854, United States. Electronic address: keuhrich@rutgers.edu.
authors.forename: Kathryn E
authors.initials: KE
authors.lastname: Uhrich
mesh_terms: D002412:Cations; D045744:Cell Line, Tumor; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry*; D020868:Gene Silencing; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D046911:Macromolecular Substances; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry
chemical_list: D002412:Cations; D005229:Fatty Acids, Monounsaturated; D046911:Macromolecular Substances; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D034741:RNA, Small Interfering; C020888:1,2-dielaidoylphosphatidylethanolamine; D008156:Luciferases; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane
keywords: Cationic amphiphilic macromolecule; Lipid; siRNA delivery
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