jsmc-10018

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

Beston F. Nore*

* Department of Biochemistry, School of Medicine, Faculty of Medical Sciences, University of Sulaimani, Sulaimani, Kurdistan Region, Federal Republic of IRAQ. 

Submitted: 2/10/2011; Accepted: 14/1/2012; Published 1/12/2012

DOI Link: https://doi.org/10.17656/jsmc.10018 

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. 

KEYWORDS

HIV-TAT protein; Protamine ;Transfection; Gene delivery; Cell penetrating peptides (CPPs).

References 

1. Langel, Ü. In: Cell-penetrating peptides: Methods and Protocols. Humana Press, New York, Springer Science+Business Media; 2011.

2. Porteus MH, Connelly JP, and Pruett SM. A look to future directions in gene therapy research for monogenic diseases. PLoS Genet. 2006; 2: 1285-1292.

3. Giacca, M. In: Gene Therapy. Milan: Springer-Verlag Italia; 2010.

4. Oliveira NA, Cecchi CR, Higuti E, Oliveira JE, Jensen TG, Bartolini P, and Peroni CN. Long-term human growth hormone expression and partial phenotypic correction by plasmid-based gene therapy in an animal model of isolated growth hormone deficiency. J Gene Med. 2010; 12: 580-585.

5. Bolhassani A. Potential efficacy of cell-penetrating peptides for nucleic acid and drug delivery in cancer. Biochim Biophys Acta. 2011; 1816: 232-246.

6. Choi YS, Lee JY, Suh JS, Lee SJ, Yang VC, Chung CP, and Park YJ. Cell penetrating peptides for tumor targeting. Curr. Pharm. Biotechnol. 2011; 12: 1166-1182.

7. Thorén PE, Persson D, Esbjörner EK, Goksör M, Lincoln P, Nordén B. Membrane binding and translocation of cell-penetrating peptides. Biochemistry. 2004; 43: 3471-3489.

Full Text 

 © The Authors, published by University of Sulaimani, College of Medicine

This work is licensed under a Creative Commons Attribution 4.0 International License.