Description: Cataracts is the leading cause of blindness in the world. Oxidation of the lens crystallin protein, which leads to its misfolding and aggregation, is the main cause of cataracts. Among the 20 million people affected annually, over 50% are caused by old age. Currently, eye surgery is the only effective method for treatment. This method, however, is expensive and invasive. Therefore, our goal is to prevent and treat age related cataracts non-invasively by producing an eyedrop that includes glutathione reductase (GSR) for prevention and Cholesterol 25-Hydroxylase (CH25H) for treatment. GSR catalyzes the conversion from Glutathione disulfide (GSSG) to glutathione (GSH) in order to prevent crystallin oxidation. CH25H converts cholesterol into 25-hydroxycholesterol, which is a molecule that can reverse aggregation and restore solubility of the crystallin protein. To deliver proteins into the lens more efficiently, we engineered an eyedrop containing biodegradable Chitosan nanoparticles that encapsulate the proteins.Currently, we are working on building our Biobrick constructs. In order to prevent direct contact between human body and bacteria, we assembled polyhistidine tag behind each gene we want to express to purify proteins. Furthermore, we used a strong promoter and strong ribosome to maximize protein expression. To test if GSR prevents and CH25H treats cataracts, we simulate healthy lenses and cataract lenses. We extracted the lens protein of a type of freshwater fish and dissolved the protein in Tris/DTT/EDTA buffer, which mimics the environment of the fish lens. We added GSH to test for the effectiveness of the antioxidant. For treatment, we added 25-hydroxycholesterol after adding hydrogen peroxide into the lens protein solution for 24 hours. We measured the opacity of the solution by measuring the absorbance using a spectrophotometer, which reflects the severity of cataracts condition.While we strive to develop an accurate experimental model to test our methods of prevention and treatment of cataracts, we have also taken various measures to further our research, educate the public, and fundraise for cataract patients. Through interviews with ophthalmologists and veterinarians, we gained insight into the specific requirements of an efficient solution for cataracts. In order to spread awareness of cataracts and synthetic biology as an emerging field of study, we presented our project to other students and taught introductory biology classes about the basics of synthetic biology. Finally, we conducted surveys and interviews to learn more about the opinions of the people in Taipei about the issue of cataracts and cataract surgery.
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Year: 2016Visit Wiki
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Updated at: 8/9/16