Light-Triggerable Liposomes for Enhanced Endolysosomal Escape and Gene Silencing in PC12 Cells.

Chen, W; Deng, W; Goldys, EM

Light-Triggerable Liposomes for Enhanced Endolysosomal Escape and Gene Silencing in PC12 Cells.

Chen, W Deng, W

Goldys, EM

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Publisher:: Nature Publishing Group
Publication Type:: Journal Article
Citation:: Molecular Therapy: Nucleic Acids, 2017, 7, pp. 366-377
Issue Date:: 2017-06-16

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Field	Value	Language
dc.contributor.author	Chen, W
dc.contributor.author	Deng, W https://orcid.org/0000-0002-9413-0978
dc.contributor.author	Goldys, EM
dc.date.accessioned	2022-09-06T00:18:28Z
dc.date.available	2017-04-18
dc.date.available	2022-09-06T00:18:28Z
dc.date.issued	2017-06-16
dc.identifier.citation	Molecular Therapy: Nucleic Acids, 2017, 7, pp. 366-377
dc.identifier.issn	2162-2531
dc.identifier.issn	2162-2531
dc.identifier.uri	http://hdl.handle.net/10453/161395
dc.description.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.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	Nature Publishing Group
dc.relation	http://purl.org/au-research/grants/arc/DE130100894
dc.relation	http://purl.org/au-research/grants/arc/CE140100003
dc.relation.ispartof	Molecular Therapy: Nucleic Acids
dc.relation.isbasedon	10.1016/j.omtn.2017.04.015
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0601 Biochemistry and Cell Biology, 1103 Clinical Sciences
dc.title	Light-Triggerable Liposomes for Enhanced Endolysosomal Escape and Gene Silencing in PC12 Cells.
dc.type	Journal Article
utslib.citation.volume	7
utslib.location.activity	United States
utslib.for	0601 Biochemistry and Cell Biology
utslib.for	1103 Clinical Sciences
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
utslib.copyright.status	closed_access	*
pubs.consider-herdc	false
dc.date.updated	2022-09-06T00:18:23Z
pubs.publication-status	Published
pubs.volume	7

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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http://hdl.handle.net/10453/161395