Lama2 lentiviral payload. Directing a treatment for muscular dystrophy

Abstract

In this degree final project will be summarized the results from one of the experimental procedures performed in the Translational Synthetic Biology Department in the PRBB-UPF, a research group which has a project focused on finding a treatment for merosin-deficient congenital muscular dystrophy (MCD1A) disease, a variation of congenital muscular dystrophy (CMD) which may be caused by mutations in LAMA2 gene. These mutations may provoke a deficiency or a functional reduction of laminin α2 subunit, the heavy chain of Laminin-211 molecule, also known as merosin, a cell-adhesion molecule that is highly expressed in the basement membrane on skeletal muscle and protects the muscle fiber from damage because of the constant stress of muscle contractions. We aim to explore a curative treatment for Lama2 patients using animal models to explore its efficacy. To achieve this purpose, we will improve the expression of mice Lama2 gene in bone marrow cells by using lentiviral vectors to treat muscular dystrophy. To carry out this process we cloned Lama2 gene into lentiviral vector for this way infect mammal cells with lentivirus and, by extracting RNA, DNA and proteins from those cells, check the lentiviral titer with quantitative PCR assay to analyse its infective capacity. The results of the project have shown that despite the huge length Lama2 gene it’s possible to clone into lentiviral vectors and then by lentivirus infection, transfer the transgene to mammalian cells so that it produces functional proteins in a ratio that can be used to treat the disease.