Generation and Characterization of Dysferlin Deficient and Myoferlin Deficient Muscle Cells
Original project description:
This project is characterizing existing dysferlin deficient muscle cell lines and if necessary generating further C2C12 cell line deficient for dysferlin. These cell lines and those deficient for myoferlin will then be used to generate further cell lines in which the DysF containing ferlins (dysferlin, myoferlin, Fer1L5) are simultaneously knocked down. The specific objectives are to 1) Evaluate the quality of the various commercially available dysferlin antibodies by Western blotting, immunofluorescence and immunohistochemistry. Muscle cell lines, primary muscle cultures and muscle sections will be tested. 2) Characterize the gene and protein expression of dysferlin, myoferlin and FER1L5 in the various dysferlin deficient muscle cells lines and control cells. Proliferating and differentiating cells will be analyzed. 3) Examine the expression levels of myoferlin in the AJ dysferlin deficient cells and proceed to knockdown myoferlin in these cells to evaluate their membrane resealing capability. Alongside this dysferlin will be knocked down in the myoferlin deficient C2C12 cell lines generated by our group. 4) Generate and characterize C2C12 clones deficient for dysferlin and those in which myoferlin is also knocked down.
This project generated a number of different cell lines deficient for dysferlin, myoferlin, or both. Click here to get information about how you can obtain these cells lines.