Polylysine/DNA

Zauner W, Kichler A, Schmidt W, Sinski A, Wagner E (1996) Glycerol enhancement of ligand-polylysine/DNA transfection. Biotechniques 20 905-913... 

Wagner E, Zatloukal K, Cotten M, Kirlappos H, Mechtler K, Curiel DT, Bimstiel ML (1992) Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-... 

In common with the polylysine DNA-condensing peptide, the p peptide has also been shown to have nuclear localization properties. Confocal microscopy revealed that p peptide in a complex containing fluorescent lipid- and dye-labelled DNA associates with the nuclei and nucleoli of both dividing and nondividing cells within 15 minutes of exposure to the complex, thus suggesting an NLS function. However, this property was not detectable when the peptide became incorporated into a 3p-[N-(N, N -dimethylaminoethane)-carbamoyl]-cholesterol (DC-chol)/DOPE cationic liposome (149). It may be the case that the lipids may mask the critical residues. 

Wagner E, Plank C, Zatloukal K, et al. Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin-polylysine-DNA complexes toward a synthetic virus-like gene-transfer vehicle. Proc Natl Acad Sci USA 1992 89(17) 7934-7938. 

There is a wide variety of vectors used to deliver DNA or oligonucleotides into mammalian cells, either in vitro or in vivo. The most common vector systems are based on viral [retroviruses (9, 10), adeno-associated virus (AAV) (11), adenovirus (12, 13), herpes simplex virus (HSV) (14)] andnonviral [cationic liposomes (15,16), polymers and receptor-mediated polylysine-DNA] complexes (17). Other viral vectors that are currently under development are based on lentiviruses (18), human cytomegalovirus (CMV) (19), Epstein-Barr virus (EBV) (20), poxviruses (21), negative-strand RNA viruses (influenza virus), alphaviruses and herpesvirus saimiri (22). Also a hybrid adenoviral/retroviral vector has successfully been used for in vivo gene transduction (23). A simplified schematic representation of basic human gene therapy methods is described in Figure 13.1. 

Curiel, D.T. (1994). High-efficiency gene transfer employing adenovirus-polylysine-DNA complexes. Nat. Immun., 13, 141-164. 

Erbacher, P., Roche, A.C., Monsigny, M. and Midoux, P. (1995) Glycosylated polylysine/DNA complexes Gene transfer efficiency in relation with the size and the sugar substitution level of glycosylated polylysines and with the plasmid size. Bioconjug. Chem., 6, 401—410. 

McKenzie, D.L., Collard, W.T. and Rice, K.G. (1999) Comparative gene transfer efficiency of low molecular weight polylysine DNA-condensing peptides. J. Pept. Res., 54, 311-318. 

Mislick, K. A., Baldeschwieler, J. D., Kayyem, J. F., and Meade, T. J. Transfection of folate-polylysine DNA complexes Evidence for lysosomal delivery. Bioconjug. Chem. 6 512-515, 1995. 

Wagner, E., Zatloukal, K., Cotten, M., Kirlappos, H., Mechtler, K., Curiel, D.T., and Bimstiel, M.L. (1992). Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes. Proc. Natl. Acad Sci. USA 89, 6099 6103. 

Lysosomotropic agents, such as chloroquine and sucrose, have been used to enhance DNA/vector release into the cytoplasm in vitro [187]. Viral DNA vectors have evolved the ability to escape acidified endosomes. To mimic this property, replication-defective adenoviruses were administered with transferrin-polylysine DNA vectors, resulting in greater gene expression [188,189]. More recently, endosomolytic structures have been attached to DNA vectors in lieu of using whole viruses. HA-2, the fusogenic peptide of the influenza virus... 

Stankovics, J., Crane, A. M Andrews, E Wu, C. H., Wu, G. Y., andLedley, F. D. (1994) Overexpression of human methylmalonyl CoA mutase in mice after in vivo gene transfer with asialoglycoprotein/polylysine/DNA complexes. Hum. Gene Ther. 5(9), 1095-1104. 

Erbacher P, et al. (1997). The reduction of the positive charges of polylysine by partial gluconoylation increases the transfection efficiency of polylysine/DNA complexes. Biochim. Biophys. Acta. 1324 27-36. 

Transfemn/po/y/ys/ne/DA/A—hematopoietic cells, T-cells, pulmonary epithelial cells. Asialoorosomucoid/polylysine/DNA—fNerceWs. 

Several small molecules, Including folate, galactose, lactose, and N-acetyl galactosamlne, also have been used to prepare polylysIne/DNA complexes for delivery of DNA by receptor-mediated endocytosis. 

The DNA is rapidly eliminated from the compartment into which it is administered as a result of both degradation through endo- and exonucleases and distribution to other compartments. The DNA is eliminated from the blood compartment by interaction with Kuppfer cells through a specific scavenger/ receptor interaction (173). An intravenously administered asialo-orosomucoid-polylysine-DNA complex is cleared from the blood with an apparent half-life of 2.5 min (174). The DNA taken up by the liver is eliminated with a half life of 1.0 to 1.3 hours, gene transcripts were evident for 1 to 12 hours, and the gene product persisted for 6 to 24 hours. 

Gao L, Wagner E, Gotten M, Agarwal S, Harris C, Romer M, et al. Direct in vivo gene transfer to airway epithelium employing adenoviras-polylysine-DNA complexes. Hum Gene Ther 1993 4 17-24. 

It has become very clear that virases have evolved in such a way that these agents possess the ability to circumvent the endosomal shunt. In that context, adenoviros has been complexed to polylysine- DNA to create a ternary complex that allows for more efficiency in transgene expression. It is thought that adenoviroses are able to protect their genomic DNA due to the exposure of the hy-... 

The two classes of non-viral vector that are being investigated most actively are the cationic lipid-based vectors and the cationic polymer-based vectors. The cationic polymer-based vectors represent the newest class of non-viral delivery system to be developed and there are a number of groups working with these vectors. As this is such a new area of research there has yet to be any standardised definition of the terms involved in this field. Complexes formed between cationic polymers and DNA have been referred to as interpolyelectrolj1 e complexes, molecular conjugates, polylysine-DNA complexes, DNA-polylysine complexes, polyplexes and so on. In this review complexes formed between poly-(L-lysine) (pLL) and DNA are referred to as polylysine/DNA complexes or abbreviated to pLL/DNA complexes. Where polymers other dian poly(L-lysine) have been used the term polymer/DNA complex is used. The forward slash (/) is used to denote a non-covalent interaction, while a hyphen (-) is used to denote a covalent link. 

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