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Optimization transgene expression of short fully synthesized lipopeptide-based transfection agent for non-viral gene delivery vehicle
Tarwadia,b, Restiani D.E.a, Pambudi S.a, Jazayeri J.J.c, Pouton C.W., Rachmawati H.b, Kartasasmita R.E.b, Asyarie S.b
a Centre for Pharmaceutical and Medical Technology, Agency for the Assessment and Application of Technology (BPPT), Jakarta, 10340, Indonesia
b School of Pharmacy, Bandung Institute of Technology, West Java, 40132, Indonesia
c School of Biomedical Science, Faculty of Science-Charles Sturt University, Wagga-Wagga, 2678, Australia
d Victoria College of Pharmacy- Monash University, Parkville, 3052, Australia
[vc_row][vc_column][vc_row_inner][vc_column_inner][vc_separator css=”.vc_custom_1624529070653{padding-top: 30px !important;padding-bottom: 30px !important;}”][/vc_column_inner][/vc_row_inner][vc_row_inner layout=”boxed”][vc_column_inner width=”3/4″ css=”.vc_custom_1624695412187{border-right-width: 1px !important;border-right-color: #dddddd !important;border-right-style: solid !important;border-radius: 1px !important;}”][vc_empty_space][megatron_heading title=”Abstract” size=”size-sm” text_align=”text-left”][vc_column_text]© 2018 Pharmainfo Publications. All rights reserved.Gene therapy using non-viral gene delivery vehicle might be applied for the treatment of viral infection, cancer and genetic disorders. However, the most challenging barrier is the low efficiency of transgene expression. Therefore, optimization of transgene expression is very important and critical to be performed before doing extensive evaluation of the transfection agent. Herein, we present transgene expression optimization data of gene encoding luciferase condensed with a series of short fully synthesized lipopeptide on COS7, HeLa and 293T cells. The complex formation, effect of molar ratio, forced sedimentation, incubation period of the DNA-transfection agent complexes were explored on transfection efficiency. Moreover, the enhancement effect in transfection efficiency of polymer PEI as co-transfection agent in DNA-lipopeptide was also investigated. The results show that the complex particles of the DNA-lipopeptide were formed efficiently in the low ionic strength environment (HEPES Glucose Buffer pH 7.4), and the complex was incubated more than 10 hours before it was transfected onto the cells. Depending on the lipopeptide, the DNA-lipopeptide incubation in 24 hours (at 4-6oC) resulted an increased up to 2-4-fold in the transgene expression. In addition, polymer PEI enhanced the transfection efficiency of the lipopeptide-mediated gene delivery up to 50-fold compared to control. In summary, short and a linear of fully synthesized lipopeptide-based transfection agent facilitated transgene expression in several mammalian cells by optimizing the DNA-lipopeptide complex formation and transfection condition.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Author keywords” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Indexed keywords” size=”size-sm” text_align=”text-left”][vc_column_text]Complex formation,Lipopeptide,Transfection,Transgene expression[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Funding details” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”DOI” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][/vc_column_inner][vc_column_inner width=”1/4″][vc_column_text]Widget Plumx[/vc_column_text][/vc_column_inner][/vc_row_inner][/vc_column][/vc_row][vc_row][vc_column][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][/vc_column][/vc_row]