Light-triggerable liposomes for enhanced endolysosomal escape and gene silencing in PC12 cells

Wenjie Chen, Wei Deng, Ewa M. Goldys

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Liposomes are an effective gene and/or drug delivery system, widely used in biomedical applications including gene therapy and chemotherapy. Here, we designed a photo-responsive liposome (lipVP) loaded with a photosensitizer verteporfin (VP). This photosensitizer is clinically approved for photodynamic therapy (PDT). LipVP was employed as a DNA carrier for pituitary adenylyl cyclase-activating polypeptide (PACAP) receptor 1 (PAC1R) gene knockdown in PC12 cells. This has been done by incorporating PAC1R antisense oligonucleotides inside the lipVP cavity. Cells that have taken up the lipVP were exposed to light from a UV light source. As a result of this exposure, reactive oxygen species (ROS) were generated from VP, destabilizing the endolysosomal membranes and enhancing the liposomal release of antisense DNA into the cytoplasm. Endolysosomal escape of DNA was documented at different time points based on quantitative analysis of colocalization between fluorescently labeled DNA and endosomes and lysosomes. The released antisense oligonucleotides were found to silence PAC1R mRNA. The efficiency of this photo-induced gene silencing was demonstrated by a 74% ± 5% decrease in PAC1R fluorescence intensity. Following the light-induced DNA transfer into cells, cell differentiation with exposure to two kinds of PACAP peptides was observed to determine the cell phenotypic change after PAC1R gene knockdown.

LanguageEnglish
Pages366-377
Number of pages12
JournalMolecular Therapy - Nucleic Acids
Volume7
DOIs
Publication statusPublished - 16 Jun 2017

Fingerprint

PC12 Cells
Gene Silencing
Liposomes
Gene Knockdown Techniques
Light
Photosensitizing Agents
Antisense Oligonucleotides
DNA
Adenylyl Cyclases
Peptides
Antisense DNA
Gene Transfer Techniques
Endosomes
Photochemotherapy
Drug Delivery Systems
Ultraviolet Rays
Lysosomes
Genetic Therapy
Cell Differentiation
Reactive Oxygen Species

Bibliographical note

Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • endosomal escape
  • gene delivery
  • light-responsive
  • liposomes
  • verteporfin

Cite this

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abstract = "Liposomes are an effective gene and/or drug delivery system, widely used in biomedical applications including gene therapy and chemotherapy. Here, we designed a photo-responsive liposome (lipVP) loaded with a photosensitizer verteporfin (VP). This photosensitizer is clinically approved for photodynamic therapy (PDT). LipVP was employed as a DNA carrier for pituitary adenylyl cyclase-activating polypeptide (PACAP) receptor 1 (PAC1R) gene knockdown in PC12 cells. This has been done by incorporating PAC1R antisense oligonucleotides inside the lipVP cavity. Cells that have taken up the lipVP were exposed to light from a UV light source. As a result of this exposure, reactive oxygen species (ROS) were generated from VP, destabilizing the endolysosomal membranes and enhancing the liposomal release of antisense DNA into the cytoplasm. Endolysosomal escape of DNA was documented at different time points based on quantitative analysis of colocalization between fluorescently labeled DNA and endosomes and lysosomes. The released antisense oligonucleotides were found to silence PAC1R mRNA. The efficiency of this photo-induced gene silencing was demonstrated by a 74{\%} ± 5{\%} decrease in PAC1R fluorescence intensity. Following the light-induced DNA transfer into cells, cell differentiation with exposure to two kinds of PACAP peptides was observed to determine the cell phenotypic change after PAC1R gene knockdown.",
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Light-triggerable liposomes for enhanced endolysosomal escape and gene silencing in PC12 cells. / Chen, Wenjie; Deng, Wei; Goldys, Ewa M.

In: Molecular Therapy - Nucleic Acids, Vol. 7, 16.06.2017, p. 366-377.

Research output: Contribution to journalArticleResearchpeer-review

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