*5.1.2.2. RNA electroporation*

Transfection Efficiency=KNT2.3(1 - EP / E)f(ADN) (15)

where plasmid concentration f(AND) is complex and high level of plasmid is toxic [129] and K is constant. As results, for DNA electrotransfer, the pulse effect (Field strength, short high amplitude pulse, long low amplitude pulse) are very important and which is the major

The electroporation technique has been used widely for transfection of plasmid in vitro and in vivo. Recently this technique has been used for application of DNA vaccine and gene therapies for clinical trials. Electroporation technology are not only the basis for human studies, but also it influence veterinary medical for animals, which can make the bridge between human and animal studies [130-134]. In this section, different clinical trials with electroporation

DNA vaccines have excellent potential as preventive or therapeutic agents against can‐ cers and infectious diseases. For a successful DNA delivery into the cell or tissues, DNA must need to subsequently achieve gene expression of the encoded protein at desired level or for the desired duration of time. In vivo electroporation, which can enhance the delivery efficiency and the cellular uptake of an agent by 1,000 times and it can increase the levels of gene expression (i.e. production of the coded protein) by 100 times or more compared to plasmid DNA delivered without other delivery enhancements. DNA vaccination by electroporation technique has been developed in last several years [134-140]. For DNA vaccination by electroporation, preclinical trials for mouse studies revealed that xenogene‐ ic DNA vaccination with gene encoding tyrosinase family membrane can induced anti‐ body and cytotoxic T cell responses resulted in tumor rejection [141-142]. DNA vaccine, p.DOM-PSMA encoded a domain (DOM) of fragment C of tetanus toxin to induced CD4+ T cell helps to fuse to a tumor-derived epitope from prostate-specific membrane antigen (PSMA) for use in HLA-A2+ patients with recurrent prostate cancer [139]. For this open level phase I/II work, DNA was delivered by intracellular injection followed by electropo‐ ration with five patients per dose level. Plasmid DNA vaccination using electroporation able to elicited robust humoral and CD8+ T-cell immune responses, while limited invasive‐ ness of delivery [140]. DNA delivered method which included phase I clinical trial investigated safety and immunogenicity of xenogenic tyrosinase DNA vaccine, adminis‐ tered intramuscularly with electroporation to patient with stage IIB, IIC,III or IV melano‐ ma(Clinical Trials. Gov ID NCT00471133). Electroporation with xenogeneic tyrosinase DNA vaccine can increase the human response and anti-tumor effects compared to the vaccine

parameters for efficient gene expression into cell and tissues.

80 Advances in Micro/Nano Electromechanical Systems and Fabrication Technologies

*5.1.2. Electrotransfer for clinical developments*

techniques are mentiond below.

*5.1.2.1. DNA vaccine*

alone [143].

The RNA transfer by electroporation technique has been increases continuously recently. RNA can enter inside the cell alone or be used for transfection of dendritic cells, showing several advantages as a vaccine including feasibility, applicability, safeness, and effectiveness, when it comes to the generation of immune responses. In vitro experiment, dendritic cells (DC) pulsed with whole tumor RNA or RNA encoding specific antigen like TAAs induced the generation of specific positive cytotoxic T lymphocytes (CTLs) into the cell [144]. Electropo‐ rated monocyte-derived DCs with whole RNA from LP-1, U266 cell lines and induced specific CTLs that lysed LP-1 and U266 myeloma cells [145]. The RNA delivery into the dendritic cells (DCs) can be achieved by using electroporation of dendritic cells in the presence of RNA [146]. In vivo study was performed by TriMix dendritic cells (DCs) [146]. The transfection of Dendritic cells (DCs) were performed by electroporation technique with mRNA encoding CD40L, CD70, and a constitutively actived TLR4 as enhancing elements. Additionally the cells were electroporated with either Mage-A3, Mage-C2, tyrosinase, or gp100 mRNA. The intra‐ dermal injections at four different sites of 1.25 × 107 TriMix DCs per antigen were provided to two melanoma patient in four times per week. While antigen-specific CD8 T cell responses was detected in both patients after finished treatment, but no data published for this trials [147].
