I have a mouse model that mimics Type I diabetes in that the mouse\'s pancreas d
ID: 261145 • Letter: I
Question
I have a mouse model that mimics Type I diabetes in that the mouse's pancreas does not make any insulin. Insulin is the hormone that causes cells to take up glucose from the blood thereby lowering blood glucose levels. I am trying to develop a treatment by inserting the human insulin gene into the mouse using a retrovirus that inserts itself into liver cells. Basically, I have a retrovirus with and without the human insulin gene cloned into it. (A.) Describe a possible control for this experiment to prove that a retrovirus by itself does not e an effect on blood glucose levels (5 pts). (B.) I am going to perform a Southern Blot to determine whether the virus (with the insulin gene) has inserted into the DNA of cells. Which tissue should I examine and why am ooking at this tissue? (5 pts) (C.) How can I distinguish between the human insulin gene and the mouse insulin gene that is already in the animal? (5 pts) (D.) I performed my Southern Blot using the tissue you described in B and the method of detecting the human insulin gene you described in C and got the following results. Which of the animals should I use for further study to determine if this treatment might be useful for controlling blood glucose levels and why did you choose these animals? (5 pts) -con con 2Explanation / Answer
(A)
The concentration of blood glucose decreased still was above the normal range between 2 and 8 days after transfection with pLNCP2 and pLNCI2, the concentration of blood glucose in pLNCP2 rats was always lower than in pLNCI2 mouse.
It should be emphasized that, the release of insulin by the insulin gene transferred did not regulate by glucose.
pLNCP2 and pLNCI2 plasmids, inserted insulin gene respectively, were compared with pLNCX2 plasmid.
(B) The constructed viral-insulin gene was transferred into mouse fibroblast cell.
The insulin concentration in 3- day cultured mouse fibroblast cells.
Insulin gene transferred and expressed in non-pancreatic cells as a means for the treatment of diabetes was developed rapidly in the expanding gene therapy.
Retrovirus, lentivirus, adenovirus, adenoassociated virus and herpes simplex had been used as viral vectors, and the constructed viral-insulin gene was successfully transferred into diabetic mouse cells.
A gene, containing promoter, enhancer and rat type I insulin gene (a-chain, b-chain and signal peptide), was constructed into a retrovirus vector.
(C) The constructed viral-insulin gene was transferred into mouse pancreas, and the blood glucose concentrations were monitored to determine the effect of gene therapy in diabetic mouse induced by injection.
(D) Rat is use for further study.
The results in our study suggested that the activity of rat insulin enhancer/promoter gene was an important determinant for the expression of insulin gene.
From blood glucose concentration of diabetic rats, with the retroviral vector transfection of the insulin gene into pancreas, in vivo, could demonstrate this gene therapy was effective.
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