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Gene Therapy

Gene Therapy

Gene therapy is a technique where a gene is inserted into a person’s cells to prevent or treat a serious disease. The added gene may be a healthy version of the gene that is mutated in the patient, or it could be a different gene that inactivates or compensates for the patient’s disease-causing gene. For example, in patients with severe combined immunodeficiency (SCID) due to a mutation in the gene for the enzyme adenosine deaminase, a functioning version of the gene can be inserted. The...

Methods of Nuclear Reprogramming

Methods of Nuclear Reprogramming

Nuclear reprogramming is a process of transforming one cell type into an unrelated cell type by epigenetic changes that alter the cell’s original gene expression pattern. Such epigenetic changes force cells to express a different set of genes, which play a significant role in inducing transformation into other cell types. Nuclear reprogramming offers applications in reproductive cloning for livestock propagation and regenerative medicine — developing patient-specific cells for injury repair.

In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Development of an Economical DNA Delivery System by "Acufection" and its Application to Skin Research

Development of an Economical DNA Delivery System by "Acufection" and its Application to Skin Research

Published on: April 19, 2017

Nucleofection: a new method for cutaneous gene transfer?

Frank Jacobsen¹, Janine Mertens-Rill, Juergen Beller

¹Department of Plastic Surgery, BG University Hospital Bergmannsheil, Ruhr University Bochum, Bochum 44789, Germany.

Journal of Biomedicine & Biotechnology

|May 10, 2007

Summary

Nucleofection offers higher and earlier transgene expression in primary cells compared to FuGENE6. However, this method increases cytotoxicity, impacting cell proliferation and morphology.

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Area of Science:

Cell Biology
Molecular Biology
Gene Transfer Technologies

Background:

Nonviral gene transfer in primary epithelial cells faces limited transfection efficacy.
Comparing nucleofection and FuGENE6 for gene delivery in primary cells is crucial.

Purpose of the Study:

To evaluate and compare the transfection efficacy of nucleofection versus FuGENE6 in primary human cells.
To assess reporter gene expression, cell viability, and morphology following different transfection methods.

Main Methods:

Primary human keratinocytes (HKC), fibroblasts (HFB), and HaCaT cells were transfected using FuGENE6 or Amaxa Nucleofector.
Assessed beta-galactosidase expression, cell proliferation (MTT), transduction efficiency (X-gal), morphology, and cytotoxicity (CASY).

Main Results:

Nucleofection demonstrated significantly higher and earlier transgene expression (3h vs 24h) in HKC and HFB compared to FuGENE6.
FuGENE6 maintained high cell proliferation (80-90%) and normal morphology.
Nucleofection resulted in reduced keratinocyte size and lower proliferation rates (20-42%).

Conclusions:

Nucleofection achieves superior transgene expression timing and levels over FuGENE6.
Increased cytotoxicity associated with nucleofection affects cell proliferation and morphology.