Transfection of Folate-Polylysine DNA Complexes: Evidence for Lysosomal Delivery

Kimberly A. Mislick; John D. Baldeschwieler; Jon F. Kayyem; Thomas J. Meade

doi:10.1021/bc00035a002

Transfection of Folate-Polylysine DNA Complexes: Evidence for Lysosomal Delivery

Kimberly A. Mislick, John D. Baldeschwieler^*, Jon F. Kayyem, Thomas J. Meade

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

134 Scopus citations

Abstract

We are utilizing the folate receptor for the intracellular delivery of DNA. In this study, a folate-poly-L-lysine (FPLL) conjugate was synthesized and equilibrated with plasmid DNA encoding the firefly luciferase gene. The FPLL-DNA complexes were added to KB cells treated with chloroquine. Luciferase activity of cells incubated with FPLL-DNA was 6-fold higher than of cells exposed to poly-L-lysine (PLL)-DNA. The addition of free folic acid competitively inhibited the enhancement of gene expression. Removal of chloroquine from the media significantly inhibited transfection efficiency of FPLL-DNA complexes. We conclude that FPLL-DNA complexes are delivered into KB cells via folate receptor-mediated endocytosis and likely follow a lysosomal pathway into the cytoplasm.

Original language	English (US)
Pages (from-to)	512-515
Number of pages	4
Journal	Bioconjugate Chemistry
Volume	6
Issue number	5
DOIs	https://doi.org/10.1021/bc00035a002
State	Published - 1995

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biomedical Engineering
Pharmacology
Pharmaceutical Science
Organic Chemistry

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title = "Transfection of Folate-Polylysine DNA Complexes: Evidence for Lysosomal Delivery",

abstract = "We are utilizing the folate receptor for the intracellular delivery of DNA. In this study, a folate-poly-L-lysine (FPLL) conjugate was synthesized and equilibrated with plasmid DNA encoding the firefly luciferase gene. The FPLL-DNA complexes were added to KB cells treated with chloroquine. Luciferase activity of cells incubated with FPLL-DNA was 6-fold higher than of cells exposed to poly-L-lysine (PLL)-DNA. The addition of free folic acid competitively inhibited the enhancement of gene expression. Removal of chloroquine from the media significantly inhibited transfection efficiency of FPLL-DNA complexes. We conclude that FPLL-DNA complexes are delivered into KB cells via folate receptor-mediated endocytosis and likely follow a lysosomal pathway into the cytoplasm.",

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T1 - Transfection of Folate-Polylysine DNA Complexes

T2 - Evidence for Lysosomal Delivery

AU - Mislick, Kimberly A.

AU - Baldeschwieler, John D.

AU - Kayyem, Jon F.

AU - Meade, Thomas J.

PY - 1995

Y1 - 1995

N2 - We are utilizing the folate receptor for the intracellular delivery of DNA. In this study, a folate-poly-L-lysine (FPLL) conjugate was synthesized and equilibrated with plasmid DNA encoding the firefly luciferase gene. The FPLL-DNA complexes were added to KB cells treated with chloroquine. Luciferase activity of cells incubated with FPLL-DNA was 6-fold higher than of cells exposed to poly-L-lysine (PLL)-DNA. The addition of free folic acid competitively inhibited the enhancement of gene expression. Removal of chloroquine from the media significantly inhibited transfection efficiency of FPLL-DNA complexes. We conclude that FPLL-DNA complexes are delivered into KB cells via folate receptor-mediated endocytosis and likely follow a lysosomal pathway into the cytoplasm.

AB - We are utilizing the folate receptor for the intracellular delivery of DNA. In this study, a folate-poly-L-lysine (FPLL) conjugate was synthesized and equilibrated with plasmid DNA encoding the firefly luciferase gene. The FPLL-DNA complexes were added to KB cells treated with chloroquine. Luciferase activity of cells incubated with FPLL-DNA was 6-fold higher than of cells exposed to poly-L-lysine (PLL)-DNA. The addition of free folic acid competitively inhibited the enhancement of gene expression. Removal of chloroquine from the media significantly inhibited transfection efficiency of FPLL-DNA complexes. We conclude that FPLL-DNA complexes are delivered into KB cells via folate receptor-mediated endocytosis and likely follow a lysosomal pathway into the cytoplasm.

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Transfection of Folate-Polylysine DNA Complexes: Evidence for Lysosomal Delivery

Abstract

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