Myostatin (MSTN) is a negative regulator of skeletal muscle mass. Strategies to block
myostatin signaling pathway have been extensively pursued to increase muscle mass in various disease settings including
muscular dystrophy. Here, we report a new class of
reagents based on
transcription activator-like effector nucleases (
TALENs) to disrupt
myostatin expression at the genome level. We designed a pair of MSTN
TALENs to target a highly conserved sequence in the coding region of the
myostatin gene. We demonstrate that codelivery of these MSTN
TALENs induce highly specific and efficient gene disruption in a variety of human, cattle, and mouse cells. Based upon sequence analysis, this pair of
TALENs is expected to be functional in many other mammalian species. Moreover, we demonstrate that these MSTN
TALENs can facilitate targeted integration of a mCherry expression cassette or a larger
muscular dystrophy gene (
dysferlin) expression cassette into the MSTN locus in mouse or human cells. Therefore, targeted editing of the
myostatin gene using our highly specific and efficient
TALEN pair would facilitate cell engineering, allowing potential use in translational research for cell-based
therapy.Molecular
Therapy-
Nucleic Acids (2013) 2, e112; doi:10.1038/mtna.2013.39; published online 30 July 2013.