Journal of Medical Microbiology and Infectious Diseases
Journal of Medical Microbiology and Infectious Diseases
JoMMID
Medical Sciences
http://jommid.pasteur.ac.ir
1
admin
2345-5349
2345-5330
8
10.61186/JoMMID
14
8888
13
en
jalali
1397
10
1
gregorian
2019
1
1
7
1
online
1
fulltext
en
Enhancing HIV-1 Nef Penetration into Mammalian Cells as an Antigen Candidate
Other
Other
Original article
Original article
<strong>Introduction: </strong>The human immunodeficiency virus type 1 (HIV-1) Nef regulatory protein is known as a candidate for the design of therapeutic HIV DNA and protein vaccines. One of the limitations of these vaccines is the inability of DNA and protein to pass through the cell membrane. Various delivery systems have been developed to transfer DNA and protein into cells. Cell penetrating systems such as MPG and Cylop-1 are among delivery systems, which can deliver DNA and protein cargoes into the cells, respectively. <strong>Methods: </strong>In this study, we produced the recombinant Nef protein in <em>Escherichia coli</em> expression system. Then, the formation of CPP/DNA and CPP/protein nanoparticles was confirmed by agarose gel retardation, scanning electron microscope (SEM), Zetasizer and SDS-PAGE, and their stability was evaluated against nucleases and proteases. Finally, the delivery of the nanoparticles into HEK-293T cells was assessed by fluorescent microscopy, flow cytometry, and western blotting. <strong>Results: </strong>Our data confirmed the formation of stable nanoparticles through non-covalent bonds with a diameter of less than 200 nm. Moreover, the results of fluorescence microscopy, flow cytometry, and western blotting demonstrated that these CPPs could successfully deliver the Nef protein and DNA into HEK-293T cells. <strong>Conclusion: </strong>Our results indicated that the MPG and CyLoP-1 CPPs are suitable candidates for the delivery of DNA and protein cargoes into mammalian cells, respectively.
HIV infections, Gene products nef, Cell penetrating peptides, Transfection
37
43
http://jommid.pasteur.ac.ir/browse.php?a_code=A-10-123-3&slc_lang=en&sid=1
Saba
Davoodi
10031947532846004562
10031947532846004562
No
Department of Biology, School of Basic Science, Science and Research Branch, Islamic Azad University, Tehran, Iran
Azam
Bolhassani
azam.bolhassani@yahoo.com
10031947532846004563
10031947532846004563
Yes
Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran
Seyed Mehdi
Sadat
10031947532846004564
10031947532846004564
No
Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran
Shiva
Irani
10031947532846004565
10031947532846004565
No
Department of Biology, School of Basic Science, Science and Research Branch, Islamic Azad University, Tehran, Iran