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Transfection reagents

a technology of transfection reagents and reagents, applied in biochemistry apparatus and processes, instruments, material analysis, etc., can solve problems such as difficult conventional treatment, and achieve the effect of increasing cross-sectional area and reducing the hydrophobicity of reagents

Inactive Publication Date: 2006-11-16
NORTHWESTERN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The combination of cationic lipoids with varying chain lengths significantly increases transfection efficiency in human umbilical artery endothelial cells and human dermal fibroblasts, achieving up to 15% transfection efficiency in optimal conditions, addressing the limitations of traditional cationic lipids.

Problems solved by technology

There are approximately four thousand different genetic diseases, many highly debilitating and frequently resulting in death at an early age.
Because almost all of these diseases involve a defective protein, conventional treatment is difficult.

Method used

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Examples

Experimental program
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Effect test

example 1

EDLPC / EDOPC Transfection Reagents

[0072] Experiments conducted during the development of the present invention found that attention to the hydrophobic portions of medium and long-chain cationic lipids synergistically enhance transfection. It was found that a combination of two cationic lipid derivatives with the same head group but tails of different chain lengths behave considerably differently as transfection agents than the separate molecules. For example, the combination of the dilauroyl (12 carbon chain) and the dioleoyl (18 carbon chain) homologues of O-ethylphosphatidylcholine transfected DNA into primary human umbilical artery endothelial cells (HUAECS) more than 30-fold more efficiently than either compound separately. The present invention is not limited to a particular mechanism. Indeed, an understanding of the mechanism is not necessary to practice the present invention. Nonetheless, these results suggest that the hydrophobic portions of medium and long-chain cationic li...

example 2

Transfection of Human Dermal Fibroblasts with EDOPC / EPOPC and EDOPC / EDiphytanoyl PC Transfection Reagents

[0087]FIG. 8 shows that combining EDOPC with EPOPC (one oleoyl chain, which is 18C's with one double bond, and one palmitoyl chain, which is 16C's without any double bond) shows little mixing effect in transfection of human dermal fibroblast cells in the absence of serum.

[0088]FIG. 9 shows that combining EDOPC with EDiphytanoyl PC (two phytanoyl chains, 16 carbon chains with 4 methyl branches) shows marked mixing effect in the transfection of human dermal fibroblast cells in the absence of serum.

example 3

Transfection of HUAECs with EDOPC / EPOPC, EDOPC / EDiphytanoylPC, EDOPC / SDOPC and EDOPC / EC18C10PC

[0089]FIG. 10 shows the results of transfecting HUAECs with EDOPC / EPOPC, EDOPC / EDiphytanoylPC, EDOPC / SDOPC (DOPC with an 18 carbon chain instead of an ethyl group on the phosphate oxygen), and EDOPC / EC18C10PC mixtures.

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Abstract

The present invention provides optimized transfection reagents comprising mixtures of cationiclipoids. In particular, the present invention provides DNA delivery vehicles based on identifying the optimal hydrophobicity of novel cationic phospholipid derivatives that, alone or in combination, form complexes with DNA (lipoplexes) and exhibit enhanced transfection activity.

Description

[0001] This application claims priority to U.S. Provisional Patent Application Ser. No. 60 / 508,544, filed Oct. 3, 2003, which is incorporated herein by reference in its entirety.[0002] The present invention was made, in part, under funds from the National Institutes of Health Grant No. GM 52329. The government may have certain rights in the invention.FIELD OF THE INVENTION [0003] The present invention provides optimized transfection reagents comprising mixtures of cationic lipoids. In particular, the present invention provides DNA delivery vehicles based on identifying optimal hydrophobicity of novel cationic phospholipid derivatives that, alone or in combination, form complexes with DNA (lipoplexes) and exhibit enhanced transfection activity. BACKGROUND [0004] There are approximately four thousand different genetic diseases, many highly debilitating and frequently resulting in death at an early age. Because almost all of these diseases involve a defective protein, conventional trea...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K9/127C12N15/88C12NC12N15/85
CPCC12N15/88
Inventor MACDONALD, ROBERT C.WANG, LI
Owner NORTHWESTERN UNIV
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