title: Serum insensitive, intranuclear protein delivery by the multipurpose cationic lipid SAINT-2.
abstract: Cationic liposomal compounds are widely used to introduce DNA and siRNA into viable cells, but none of these compounds are also capable of introducing proteins. Here we describe the use of a cationic amphiphilic lipid SAINT-2:DOPE for the efficient delivery of proteins into cells (profection). Labeling studies demonstrated equal delivery efficiency for protein as for DNA and siRNA. Moreover, proteins complexed with Saint-2:DOPE were successfully delivered, irrespective of the presence of serum, and the profection efficiency was not influenced by the size or the charge of the protein:cationic liposomal complex. Using beta-galactosidase as a reporter protein, enzymatic activity was detected in up to 98% of the adherent cells, up to 83% of the suspension cells and up to 70% of the primary cells after profection. A delivered antibody was detected in the cytoplasm for up to 7 days after profection. Delivery of the methyltransferase M.SssI resulted in DNA methylation, leading to a decrease in E-cadherin expression. The lipid-mediated multipurpose transport system reported here can introduce proteins into the cell with an equal delivery efficiency as for nucleotides. Delivery is irrespective of the presence of serum, and the protein can exert its function both in the cytoplasm and in the nucleus. Furthermore, DNA methylation by M.SssI delivery as a novel tool for gene silencing has potential applications in basic research and therapy.
authors: Department of Pathology and Laboratory Medicine, Section Medical Biology, University Medical Center Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands. b.t.f.van.der.gun@med.umcg.nl
authors: Bernardina T F
authors: BT
authors: van der Gun
authors: Amélie
authors: A
authors: Monami
authors: Sven
authors: S
authors: Laarmann
authors: Tamás
authors: T
authors: Raskó
authors: Krystyna
authors: K
authors: Slaska-Kiss
authors: Elmar
authors: E
authors: Weinhold
authors: Reinhold
authors: R
authors: Wasserkort
authors: Lou F M H
authors: LF
authors: de Leij
authors: Marcel H J
authors: MH
authors: Ruiters
authors: Antal
authors: A
authors: Kiss
authors: Pamela M J
authors: PM
authors: McLaughlin
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D019556:COS Cells; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002412:Cations; D002467:Cell Nucleus / Q000201:enzymology / Q000378:metabolism*; D002626:Chemistry, Pharmaceutical; D002522:Chlorocebus aethiops; D004247:DNA / Q000378:metabolism; D019175:DNA Methylation; D015257:DNA-Cytosine Methylases / Q000378:metabolism; D004337:Drug Carriers; D004339:Drug Compounding; D020868:Gene Silencing; D006801:Humans; D019169:Jurkat Cells; D015394:Molecular Structure; D010316:Particle Size; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011726:Pyridinium Compounds / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000378:metabolism; D044967:Serum / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D000906:Antibodies; D015820:Cadherins; D002412:Cations; D004337:Drug Carriers; D010714:Phosphatidylethanolamines; D011506:Proteins; D011726:Pyridinium Compounds; D034741:RNA, Small Interfering; C414306:SAINT 2; C020888:1,2-dielaidoylphosphatidylethanolamine; D004247:DNA; C081573:DNA modification methylase SssI; D015257:DNA-Cytosine Methylases; D001616:beta-Galactosidase