Receptor-mediated gene transfer in vivo

Bibliographic Details
Main Author:	Perales, Jose Carlos
Language:	English
Published:	Case Western Reserve University School of Graduate Studies / OhioLINK 1995
Subjects:	Receptor-mediated gene transfer
Online Access:	http://rave.ohiolink.edu/etdc/view?acc_num=case1057931585

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spelling	ndltd-OhioLink-oai-etd.ohiolink.edu-case10579315852021-08-03T05:31:04Z Receptor-mediated gene transfer in vivo Perales, Jose Carlos Receptor-mediated gene transfer Receptor-mediated gene transfer is an attractive method for therapeutically correcting human genetic diseases since it permits the targeting of DNA to cellular receptors in specific tissues of adult animals. The gene delivery vehicle is composed of the DNA of interest, a DNA binding moiety (i.e., poly(L-lysine)) and a ligand for the target receptor. Genes introduced by this technique have been shown to be expressed in the target tissue for varying periods. However, to be useful for gene therapy, it is critical that both the chemical properties and physical interactions of the reagents involved in the design of the DNA delivery vehicle (DNA/ligand complex) be rigorously characterized. To obtain a DNA/ligand complex suitable for in vivo gene transfer, I have developed a multi-step protocol for the interaction of poly(L-lysine) with DNA in solution. I demonstrate here that the slow increase in the concentration of poly(L-lysine) in a vortexing solution of DNA results in the formation of DNA/ligand-poly(L-lysine) particles of defined size and shape. I hypothesize that binding of poly(L-lysine) to DNA initiates condensation by forming a nucleus of condensation along the DNA length. Further addition of poly(L-lysine) results in the formation of aggregated DNA/ligand-poly(L-lysine) complexes. The second step in the formation of stable condensed DNA particles consists in equilibrating the ionic strength of the solvent to facilitate the solubilization of the condensed DNA particles. Several biochemical parameters have been studied and correlated with the functional activity of DNA/ligand-poly(L-lysine) particles in transferring genes to the liver of adult animals by receptor-mediated endocytosis. Based on these findings, a model for the interaction between the DNA and the ligand/poly(L-lysine) is proposed and the implications for the effective transfer of genes into animals in vivo discussed. The model proposed indicates a role for the polycation in both the formation of suitable particles for gene transfer, and the efficient transfer of the DNA to the nucleus of cells 1995 English text Case Western Reserve University School of Graduate Studies / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=case1057931585 http://rave.ohiolink.edu/etdc/view?acc_num=case1057931585 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection	NDLTD
language	English
sources	NDLTD
topic	Receptor-mediated gene transfer
spellingShingle	Receptor-mediated gene transfer Perales, Jose Carlos Receptor-mediated gene transfer in vivo
author	Perales, Jose Carlos
author_facet	Perales, Jose Carlos
author_sort	Perales, Jose Carlos
title	Receptor-mediated gene transfer in vivo
title_short	Receptor-mediated gene transfer in vivo
title_full	Receptor-mediated gene transfer in vivo
title_fullStr	Receptor-mediated gene transfer in vivo
title_full_unstemmed	Receptor-mediated gene transfer in vivo
title_sort	receptor-mediated gene transfer in vivo
publisher	Case Western Reserve University School of Graduate Studies / OhioLINK
publishDate	1995
url	http://rave.ohiolink.edu/etdc/view?acc_num=case1057931585
work_keys_str_mv	AT peralesjosecarlos receptormediatedgenetransferinvivo
_version_	1719421201534681088

Receptor-mediated gene transfer in vivo

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