The inability of axon regeneration in adult mammalian central nervous system (CNS) has been a major question in both fundamental and clinical neuroscience. Some regulatory mechanisms on axon regeneration have been identified but our understanding is still limited. To systematically explore novel targets, we tried to set up a genetic screening for axon elongation in vitro. By using short hairpin RNA targeting Gsk3b, we successfully knocked down the gene in cells and observed more and longer neurite outgrowth. Gsk3b knockout has previously been shown to promote axon regeneration in vivo. By using shGsk3b as a positive control and a software for automatic neurite quantification, we started to test a small proportion of shRNAs from a library. Some showed potential for increasing neu...[ Read more ]

The inability of axon regeneration in adult mammalian central nervous system (CNS) has been a major question in both fundamental and clinical neuroscience. Some regulatory mechanisms on axon regeneration have been identified but our understanding is still limited. To systematically explore novel targets, we tried to set up a genetic screening for axon elongation in vitro. By using short hairpin RNA targeting Gsk3b, we successfully knocked down the gene in cells and observed more and longer neurite outgrowth. Gsk3b knockout has previously been shown to promote axon regeneration in vivo. By using shGsk3b as a positive control and a software for automatic neurite quantification, we started to test a small proportion of shRNAs from a library. Some showed potential for increasing neurite length and future experiments are needed to verify this. Together, these data indicate that we have introduced an in vitro model for genetic screening of axon elongation.