Our proposal begins with the addition of specific ligands of the protein encoded by Cacng1 to the liposomes, in order for the liposome to be directed correctly to its target cells. This gene was chosen because it encodes membrane proteins specific to the skeletal muscle and are FDA approved drug targets. These liposomes will contain, in solution, three components: the RNA encoding the Cas-9 protein, the gRNA and a stranded DNA encoding the correct Dystrophin  gene (DMD).

 

    In vivo, the specific ligand recognizes the proteins encoded by Cacng1, causing the liposomes to fuse with the cell membranes of skeletal muscle tissue, releasing the biomolecules that are stored in their interior.

 

      Inside the cell, the RNA encoding the Cas-9 protein will be transcribed, leading to production of said protein. This, in turn, will bind to the gRNA, which is the homolog to the DNA of the the defective Dystrophin gene. During the connection of these two components, the Cas-9 will perform a knock-out (cut) in the gene.

 

     The original DNA strand of said liposomes contain, in addition to the genetic sequence which encodes Dystrophin correctly, 700 bases complementary to the pre-existing DNA in the genome surrounding the sequence to be replaced. These bases, while the DNA chain is open (due to the "unzipping" of it, perfomed by Cas-9), will bind to complementary bases immediately adjacent to the cut sides, so that the correct gene sequence of the Dystrophin snaps into place.

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