BME494 Sp2014 Dhatt

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Background & Proposed Application BACKGROUND The synthetic system modeled in the “Construction of a genetic toggle switch In Escherichia coli” uses two different states, an “on” state” and an “off” state (Gardner et all 2000). The classical system uses two repressor and two promoter pairs in which both repressors are inhibited by a different inducer, aTc and IPTG. The promoters were placed next to the repressor gene of the opposite pair. This system represents a bistable gene-regulatory network where only one promoter can be expressed at one time because the expression of one promoter repressed expression of the other promoter. The on-state of the cell was represented by placing a green fluorescent protein (GFP) transcription gene downstream of the on-state promoter. Classical Toggle Switch Design (Gardner et al, 2000) APPLICATION OF MY PROPOSED NEW DEVICE Diabetes mellitus, or diabetes, is a group of metabolic diseases in which an individual has high blood sugar. Diabetes occurs due to the either the pancreas not producing enough insulin or because cells of the body do not respond to the insulin being produced. There are three types of diabetes: Type I, Type II, and gestational diabetes. Serious long-term complications of diabetes include heart disease, kidney failure, and damage to the eyes. Diabetes is a chronic disease and there is no known preventative measure for type I diabetes. Having awareness of the disease and treating it at early stages can prevent and lower the risk of complications in patients with diabetes. Patients with diabetes must maintain keeping both short-term and long-term blood glucose levels within acceptable ranges. Globally, 227-285 million individuals have diabetes and about 90% of these individuals have Type II diabetes. In 2011, 1.4 million deaths occurred worldwide due to the result of diabetes making it the 8th leading cause of death. This number is estimated to double in the next 15 years, therefore, there is a need for a rapid diagnostic test. Design of a New Device The device will be designed as a diagnostic tool for diabetes and the functionality of the genetic switch replicates that of an “AND” logic gate. The device will require two conditions to be true in order for an output to be produced. One condition is that IPTG must be present in the device’s environment. When IPTG is present, it will bind to the LacI repressor and allow for transcription to continue. The other condition is that glucose levels in the device’s environment must be low. Glucose levels affect production of cAMP inversely; when glucose levels are high, cAMP production decreases and when glucose levels are low, cAMP production increases. cAMP binds to catabolite activator protein (CAP_ to form the CAP-cAMP complex. For the complex, cAMP must be present and glucose levels must be low. This complex is the required input of the device. In the natural lac operon, the CAP-cAMP complex leads to activation of gene expression from the lac operon. If glucose is present, cAMP levels will be low and the host will metabolize glucose if lactose is present. Building the New Device SYNTHETIC DNA LAYOUT RESOURCES TYPE IIS ASSEMBLY Testing the New Device LAC OPERON MODEL SIMULATION I used a model of the natural Lac operon to learn how changing the parameter values changes the behavior of the system. RELATIONSHIP BETWEEN THE LAC MODEL AND MY NEW DESIGN Similarities: Differences: TESTING THE NEW DEVICE Human Practices Danger of Chemicals in Farmlands About the Designer Your Name My name is ###, and I am a ### majoring in ###. I am taking BME 494 because ###. An interesting fact about me is that ###. References [1] Full reference. [2] Full reference. [3] Full reference.