Pasquet Lise, Chabot Sophie, Bellard Elisabeth, Rols Marie-Pierre, Teissie Justin, Golzio Muriel
Institut de Pharmacologie et de Biologie Structurale, Universite de Toulouse, CNRS, Toulouse, France.
Hum Gene Ther Methods. 2019 Feb;30(1):17-22. doi: 10.1089/hgtb.2018.051. Epub 2019 Jan 31.
The skin is considered as well suited for gene therapy and vaccination. DNA vaccines elicit both broad humoral and cellular immune responses when injected in the skin. Physical and chemical methods are needed to boost the expression. Gene electrotransfer (GET) is one of the most effective approaches. This step-by-step protocol describes the procedures to obtain an efficient GET targeted to the skin by using easy-to-use noninvasive electrodes after intradermal plasmid injection (i.d. GET). A specific pulse sequence is reported. Expression is observed by in vivo fluorescence imaging during >2 weeks as the plasmid was coding for tdTomato. The protocol is efficient for the transient expression of clinical proteins.
皮肤被认为非常适合基因治疗和疫苗接种。DNA疫苗注射到皮肤中时会引发广泛的体液免疫和细胞免疫反应。需要物理和化学方法来促进表达。基因电穿孔转移（GET）是最有效的方法之一。本分步方案描述了在皮内注射质粒后使用易于操作的非侵入性电极获得靶向皮肤的高效GET（皮内GET）的程序。报告了一个特定的脉冲序列。由于质粒编码tdTomato，在超过2周的时间内通过体内荧光成像观察到表达情况。该方案对于临床蛋白质的瞬时表达是有效的。
