Seeing the gene therapy: Application of gene gun technique to transfect and decolour pigmented rat skin with human agouti signalling protein cDNA

C. H. Yang, S. C. Shen, J. C. Lee, Ping-Ching Wu, S. F. Hsueh, C. Y. Lu, C. T. Meng, H. S. Hong, L. C. Yang

研究成果: Article

15 引文 (Scopus)

摘要

We developed a gene gun method for the transfer of human agouti signalling protein (ASP) cDNA to alter rat skin colour in vivo. Human ASP cDNA was cloned into a modified cytomegalovirus plasmid and delivered to the skin of Long-Evans rats by gene gun bombardment. Skin pigmentation, body weight and blood sugar of ASP cDNA-transfected rats were recorded against the control group, which were injected with plasmids encoding for green fluorescent protein. The treated skin showed lighter skin colour after 3 days of ASP gene transfection. This depigmentation effect was most prominent on day 14 and the skin gradually returned to its original pigmentation by day 28. Successful transfection of ASP gene in skin and hair follicles, as well as downregulation of melanocortin-1 receptor (MC1R) and tyrosinase expression upon treatment, was confirmed using immunohistochemistry and Western blot analysis. Body weight and blood sugar in the treated rats did not show statistically significant differences as compared to control groups. These observations demonstrate that gene transfer using the gene gun method can induce high cutaneous ASP production and facilitate a switch from dark to fair colour without systemic pleiotropic effects. Such a colour switch may be that ASP is acting in a paracrine fashion. In addition, this study verifies that ASP exerts its functions by acting as an independent ligand that downregulates the melanocyte MC1R and tyrosinase protein in an in vivo system. Our result offers new, interesting insights about the effect of ASP on pigmentation, providing a novel approach to study the molecular mechanisms underlying skin melanogenesis.

原文English
頁(從 - 到)1033-1039
頁數7
期刊Gene Therapy
11
發行號13
DOIs
出版狀態Published - 2004 七月 1

指紋

Agouti Signaling Protein
Biolistics
Genetic Therapy
Complementary DNA
Receptor, Melanocortin, Type 1
Skin
Skin Pigmentation
Firearms
Genes
Monophenol Monooxygenase
Pigmentation
Transfection
Blood Glucose
Plasmids
Down-Regulation
Color
Body Weight
Long Evans Rats
Control Groups
Hair Follicle

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

引用此文

Yang, C. H. ; Shen, S. C. ; Lee, J. C. ; Wu, Ping-Ching ; Hsueh, S. F. ; Lu, C. Y. ; Meng, C. T. ; Hong, H. S. ; Yang, L. C. / Seeing the gene therapy : Application of gene gun technique to transfect and decolour pigmented rat skin with human agouti signalling protein cDNA. 於: Gene Therapy. 2004 ; 卷 11, 編號 13. 頁 1033-1039.
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abstract = "We developed a gene gun method for the transfer of human agouti signalling protein (ASP) cDNA to alter rat skin colour in vivo. Human ASP cDNA was cloned into a modified cytomegalovirus plasmid and delivered to the skin of Long-Evans rats by gene gun bombardment. Skin pigmentation, body weight and blood sugar of ASP cDNA-transfected rats were recorded against the control group, which were injected with plasmids encoding for green fluorescent protein. The treated skin showed lighter skin colour after 3 days of ASP gene transfection. This depigmentation effect was most prominent on day 14 and the skin gradually returned to its original pigmentation by day 28. Successful transfection of ASP gene in skin and hair follicles, as well as downregulation of melanocortin-1 receptor (MC1R) and tyrosinase expression upon treatment, was confirmed using immunohistochemistry and Western blot analysis. Body weight and blood sugar in the treated rats did not show statistically significant differences as compared to control groups. These observations demonstrate that gene transfer using the gene gun method can induce high cutaneous ASP production and facilitate a switch from dark to fair colour without systemic pleiotropic effects. Such a colour switch may be that ASP is acting in a paracrine fashion. In addition, this study verifies that ASP exerts its functions by acting as an independent ligand that downregulates the melanocyte MC1R and tyrosinase protein in an in vivo system. Our result offers new, interesting insights about the effect of ASP on pigmentation, providing a novel approach to study the molecular mechanisms underlying skin melanogenesis.",
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Seeing the gene therapy : Application of gene gun technique to transfect and decolour pigmented rat skin with human agouti signalling protein cDNA. / Yang, C. H.; Shen, S. C.; Lee, J. C.; Wu, Ping-Ching; Hsueh, S. F.; Lu, C. Y.; Meng, C. T.; Hong, H. S.; Yang, L. C.

於: Gene Therapy, 卷 11, 編號 13, 01.07.2004, p. 1033-1039.

研究成果: Article

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AU - Yang, C. H.

AU - Shen, S. C.

AU - Lee, J. C.

AU - Wu, Ping-Ching

AU - Hsueh, S. F.

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AB - We developed a gene gun method for the transfer of human agouti signalling protein (ASP) cDNA to alter rat skin colour in vivo. Human ASP cDNA was cloned into a modified cytomegalovirus plasmid and delivered to the skin of Long-Evans rats by gene gun bombardment. Skin pigmentation, body weight and blood sugar of ASP cDNA-transfected rats were recorded against the control group, which were injected with plasmids encoding for green fluorescent protein. The treated skin showed lighter skin colour after 3 days of ASP gene transfection. This depigmentation effect was most prominent on day 14 and the skin gradually returned to its original pigmentation by day 28. Successful transfection of ASP gene in skin and hair follicles, as well as downregulation of melanocortin-1 receptor (MC1R) and tyrosinase expression upon treatment, was confirmed using immunohistochemistry and Western blot analysis. Body weight and blood sugar in the treated rats did not show statistically significant differences as compared to control groups. These observations demonstrate that gene transfer using the gene gun method can induce high cutaneous ASP production and facilitate a switch from dark to fair colour without systemic pleiotropic effects. Such a colour switch may be that ASP is acting in a paracrine fashion. In addition, this study verifies that ASP exerts its functions by acting as an independent ligand that downregulates the melanocyte MC1R and tyrosinase protein in an in vivo system. Our result offers new, interesting insights about the effect of ASP on pigmentation, providing a novel approach to study the molecular mechanisms underlying skin melanogenesis.

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