Crigler-Najjar type 1 disease (CN1) is a rare inherited metabolic disease characterized by complete absence of hepatic UDP-glucuronosyl transferase (UGT1), resulting in high levels of unconjugated bilirubin. CN1 is an attractive candidate disease for gene therapy. Here we show that in vivo neonatal
In vivo targeted gene transfer in skin by the use of laser-induced stress waves
✍ Scribed by Makoto Ogura; Shunichi Sato; Kuniaki Nakanishi; Maki Uenoyama; Tetsuro Kiyozumi; Daizo Saitoh; Tomosumi Ikeda; Hiroshi Ashida; Minoru Obara
- Publisher
- John Wiley and Sons
- Year
- 2004
- Tongue
- English
- Weight
- 135 KB
- Volume
- 34
- Category
- Article
- ISSN
- 0196-8092
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✦ Synopsis
Abstract
Background and Objectives
Much interest has been shown in the use of lasers for nonviral targeted gene transfer, since the spatial characteristics of laser light are quite well defined. The aim of this study was to demonstrate in vivo gene transfer by the use of laser‐induced stress waves (LISWs).
Study Design/Materials and Methods
After reporter genes had been intradermally injected to rat skin in vivo, a laser target was placed on the gene‐injected skin. LISWs were generated by the irradiation of an elastic laser target with 532‐nm nanosecond laser pulses of a Q‐switched Nd:YAG laser.
Results
Levels of luciferase activities for the skin exposed to LISWs were two orders of magnitude higher than those for the skin injected with naked DNA. Expressions of enhanced green fluorescent protein (EGFP) and β‐galactosidase were observed only in the area that was exposed to LISWs, and in addition, epidermal cells were selectively transfected. No major side effects were observed, and luciferase activity levels as high as 10^5^ RLU per mg of protein were sustained even 5 days after gene transfer.
Conclusion
Highly efficient and site‐specific gene transfer can be achieved by applying a few pulses of nanosecond pulsed LISWs to rat skin in vivo. Lasers Surg. Med. 34:242–248, 2004. © 2004 Wiley‐Liss, Inc.
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## Abstract ## Background Stability, integrity and retention of the DNA within the targeted tissue is decisive for efficient gene transfer using naked DNA. Pre‐clinical and clinical studies require reproducible transfection rates by preventing rapid degradation of naked DNA in the transduced tissu