Inclusion of TAT and NLS sequences in lipopeptide molecules generates homogenous nanoparticles for gene delivery applications.

Autor: Tarwadi; Research Center for Vaccines and Drugs, National Agency for Research and Innovation (BRIN), Building 610-611 Puspiptek Area, Tangerang Selatan, Banten 15314, Indonesia; PT Indomabs Biosantika Utama, Gedung Technology Business and Innovation Centre (TBIC), Pengasinan, Gunung Sindur, Kabupaten Bogor, Jawa Barat 16340, Indonesia. Electronic address: tarw001@brin.go.id., Pambudi S; Research Center for Vaccines and Drugs, National Agency for Research and Innovation (BRIN), Building 610-611 Puspiptek Area, Tangerang Selatan, Banten 15314, Indonesia. Electronic address: saba001@brin.go.id., Sriherwanto C; Research Centre for Applied Microbiology, National Agency for Research and Innovation (BRIN), Building 610-611 Puspiptek Area, Tangerang Selatan, Banten 15314, Indonesia. Electronic address: catu001@brin.go.id., Sasangka AN; Biomedical Sciences, Faculty of Medicine, Universitas Indonesia, Depok, Jawa Barat 16424, Indonesia. Electronic address: ayu.sasangka@sci.ui.ac.id., Bowolaksono A; Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Depok, Jawa Barat 16424, Indonesia. Electronic address: alaksono@sci.ui.ac.id., Wijayadikusumah AR; Research and Development Division, PT. Bio Farma, Jl. Pasteur No 28 Bandung, Jawa Barat 40161, Indonesia. Electronic address: acep.riza@biofarma.co.id., Zeng W; Peter Doherty Institute, The University of Melbourne, 792 Elizabeth St, Melbourne, VIC 3000, Australia. Electronic address: weiguang@unimelb.edu.au., Rachmawati H; School of Pharmacy, Bandung Institute of Technology, Jl. Ganesa 10 Bandung, Jawa Barat 40132, Indonesia; Research Centre of Nano Sciences and Nanotechnology, Bandung Institute of Technology, Jl. Ganesa 10 Bandung 40132, Jawa Barat, Indonesia. Electronic address: h_rachmawati@itb.ac.id., Kartasasmita RE; School of Pharmacy, Bandung Institute of Technology, Jl. Ganesa 10 Bandung, Jawa Barat 40132, Indonesia. Electronic address: kartasasmita@itb.ac.id., Kazi M; Department of Pharmaceutics, College of Pharmacy, POBOX-2457, King Saud University, Riyadh 11451, Saudi Arabia. Electronic address: mkazi@ksu.edu.sa.
Jazyk: angličtina
Zdroj: International journal of pharmaceutics [Int J Pharm] 2024 Sep 05; Vol. 662, pp. 124492. Date of Electronic Publication: 2024 Jul 20.
DOI: 10.1016/j.ijpharm.2024.124492
Abstrakt: Purposes: The objective of this study is to develop a versatile gene carrier based on lipopeptides capable of delivering genetic material into target cells with minimal cytotoxicity.
Methods: Two lipopeptide molecules, palmitoyl-CKKHH and palmitoyl-CKKHH-YGRKKRRQRRR-PKKKRKV, were synthesized using solid phase peptide synthesis and evaluated as transfection agents. Physicochemical characterization of the lipopeptides included a DNA shift mobility assay, particle size measurement, and transmission electron microscopy (TEM) analysis. Cytotoxicity was assessed in CHO-K1 and HepG2 cells using the MTT assay, while transfection efficiency was determined by evaluating the expression of the green fluorescent protein-encoding gene.
Results: Our findings demonstrate that the lipopeptides can bind, condense, and shield DNA from DNase degradation. The inclusion of the YGRKKRRQRRR sequence, a transcription trans activator, and the PKKKRKV sequence, a nuclear localization signal, imparts desirable properties. Lipopeptide-based TAT-NLS/DNA nanoparticles exhibited stability for up to 20 days when stored at 6-8 °C, displaying uniformity with a compact size of approximately 120 nm. Furthermore, the lipopeptides exhibited lower cytotoxicity compared to the poly-L-lysine. Transfection experiments revealed that protein expression mediated by the lipopeptide occurred at a charge ratio ranging from 4.0 to 8.0.
Conclusion: These results indicate that the lipopeptide, composed of a palmitoyl alkyl chain and TAT and NLS sequences, can efficiently condense and protect DNA, form stable and uniform nanoparticles, and exhibit promising characteristics as a potential gene carrier with minimal cytotoxicity.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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