A native function for RAN translation and CGG repeats in regulating Fragile X protein synthesis

Repeat-associated non-AUG translation of expanded CGG repeats (CGG RAN) from the FMR1 5’ UTR produces toxic proteins that contribute to neurodegeneration in Fragile X-associated Tremor/Ataxia Syndrome (FXTAS). Here we describe how unexpanded CGG repeats and their translation play conserved roles in regulating FMRP synthesis. In neurons, CGG RAN acts as an inhibitory upstream open reading frame to suppress basal FMRP production. Activation of mGluR5 receptors enhances FMRP synthesis. This enhancement requires both the CGG repeat and CGG RAN initiation sites. Using non-cleaving antisense oligonucleotides (ASOs), we selectively blocked RAN translation. This ASO blockade enhanced endogenous human neuronal FMRP expression. In human and rodent neurons, RAN blocking ASOs suppressed repeat toxicity and prolonged survival. These findings delineate a native function for CGG repeats and RAN translation in regulating basal and activity-dependent FMRP synthesis and demonstrate the therapeutic potential of modulating CGG RAN translation in fragile X-associated disorders.