Polyplex System Versus Nucleofection for Human Skin Cell Transfection and Effect of Internal Ribosome Entry Site Sequence

Tissue Engineering. Part C, Methods
Natalia Yiset Becerra ColoradoLuz Marina Restrepo Múnera

Abstract

Nonviral transfection has important implications on gene therapy because of its safety. In particular, polyfection and nucleofection are two widely used systems for nonviral gene delivery. Their potential depends on the transfection efficiency achieved, which is influenced in turn by the type of cells transfected and by the plasmid that carries the gene of interest. The efficiency of transfection by polyfection or nucleofection in human fibroblasts and keratinocytes was evaluated in this study. Transfections were performed with plasmids containing a gene of interest (human cathelicidin antimicrobial peptide) and two reporter genes (red or green fluorescent protein) that included or not an internal ribosome entry site (IRES). The efficiency was measured by flow cytometry in terms of percentage of cells expressing the reporter gene; viability of transfected cells was also evaluated. It was found that nucleofection was more efficient than polyplexes for transfecting fibroblasts, while no significant differences were found between both systems of transfection when applied to keratinocytes. Regarding the viability of fibroblasts after transfection, values were high in both systems. In contrast, keratinocytes were more sensitive to n...Continue Reading

References

Aug 10, 2000·Molecular Therapy : the Journal of the American Society of Gene Therapy·H MizuguchiT Hayakawa
May 29, 2002·Tissue Engineering·Astrid HammAnja K Bosserhoff
Jul 26, 2003·The Journal of Investigative Dermatology·Kathi C Madison
May 24, 2006·Biochimica Et Biophysica Acta·Ulrich H N DürrAyyalusamy Ramamoorthy
Feb 13, 2008·Nucleic Acids Research·Hicham BouabeJürgen Heesemann
Oct 31, 2009·Bulletin of the World Health Organization·Michael PeckDehran Swart
Mar 5, 2016·International Wound Journal·Raffaele SerraStefano de Franciscis
Jul 28, 2016·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Mariana MoralesLuz Restrepo

❮ Previous
Next ❯

Citations

Dec 7, 2019·Tissue Engineering. Part a·Maria Isabel Patiño VargasNatalia Yiset Becerra Colorado
Mar 29, 2021·Biotechnology Progress·Mirosław KucharskiEwa Ocłoń
May 14, 2021·Human Gene Therapy·Maria Isabel PatiñoPrashant K Sharma

❮ Previous
Next ❯

Datasets Mentioned

BETA
RC208872

Methods Mentioned

BETA
transfection
PCR
transfections
transfect
flow cytometry
electrophoresis
fluorescence microscopy

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.