A SYNTHETIC CELL-PENETRATING PEPTIDE (CPP) WITH PROTAMINE CONJUGATE UTILIZED FOR GENE DELIVERY

Abstract

Background

There are two main gene transfer techniques, a non-biological method such as liposome/polycations (lipofection), electroporation and gene gun. A biological method like transduction, synthetic biomolecules and virus mediated transfer. 

Objective

The objective of this study is to utilize Cell penetrating peptides (CPPs) as a novel tool for transferring and delivery of therapeutically active macromolecules and drugs in vitro and in vivo.

Materials and Methods

In this study, we utilized two synthetic CPPs, based on HIV-TAT protein, a minimal sequence of 11 amino acids with transducing domain activity. TAT is a subclass of CPPs, which often been used for intracellular targeting of proteins, but has not explored their application for DNA expression vectors. 

Results and Conclusion  

Here, we conjugated TAT to a DNA/RNA condensation moiety of Protamine, so called TAT-Protamine (TAT-P) and a branched dual TAT-Protamine (dTAT-P). Our results show that dTAT-P has both transduction capacity and DNA-condensation ability to deliver oligonucleotides and plasmid DNA into living cells. Experiments on living cells shows that there are many advantages in using dTAT-P for genetic delivery, expression plasmids of human proteins actin and Btk fusion with Green Fluorescent Protein (GFP, in addition to FITC-labelled synthetic oligonucleotides.