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Lentiviral transduction for stable collagen production and purification

a thesis in Biomedical Engineering and Biotechnology
Authors: Jacqueline Rudden Horgan;

Lentiviral transduction for stable collagen production and purification

Abstract

Burns are a major contributor to death and serious complications with an estimated 11 million cases occurring each year- to combat this, a technique was developed in Brazil utilizing the skin of a common fish, tilapia, as a replacement to the traditional harsh treatment of skin grafts. When applied as a bandage the tilapia skin helped to heal the burns effectively and quickly. Regardless of this method's functionality, there are many issues associated with this approach including ethical and sterilization concerns. To mimic this technique in a more sustainable fashion, the theory behind its functionality can be recreated using laboratory techniques. Tilapia skin contains very high amounts of type 1 collagen precursors, the most abundant type in the body. The addition of these precursors to the wound site allows the body to easily and quickly produce the additional collagen needed to repair the skin's structure. Collagen precursors can first be produced continuously in laboratory settings then included into bandages or synthetic skin as treatment for burns to help stimulate collagen production in skin cells. This project focuses on stable production of type 1 collagen through the introduction of an additional copy of the pro-α1(I) collagen precursor chain. In a laboratory setting using lentivirus transduction of a primary dermal fibroblast cell line, an additional copy of the COL1A1 gene will be added. Samples will be taken from culture media and compared against native cell lines to assess levels of collagen production. The collagen harvested will then be purified using affinity chromatography and assessed for correct conformation with use of a functional assay. The outcomes will pave the way for further advancements in the progress towards the objective of developing a viable burn treatment based on tilapia skin.

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
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