Mechanism of STMN2 cryptic splice-polyadenylation and its correction for TDP-43 proteinopathies

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JMG, SS1, Animals, Humans, Mice, DNA-Binding Proteins, Polyadenylation, RNA Precursors, Stathmin, TDP-43 Proteinopathies, RNA Splicing, RNA Splice Sites, Gene Editing, Oligonucleotides, Antisense, Neuronal Outgrowth

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Science. 2023;379(6637):1140-9.







This work was supported by NINDS/NIH R01NS112503 (to D.W.C. and C.L.-T.); NINDS/NIH RF1NS124203 (D.W.C., C.L.-T., and C.L.); ALS Finding a Cure (C.L.-T.); The Massachusetts Center for Alzheimer Therapeutic Science (C.L.-T.); The Sean M. Healey & AMG Center for ALS at Mass General (C.L.-T.); U42 Mutant Mouse Resource Research Center OD010921 (C.L.); Ruth Kirschstein Institutional National Research Service Award T32 GM008666. (M.W.B. and M.S.B.), and T32 AG 66596-2 (M.S.B.); The Packard Center for ALS Research (D.W.C. and M.W.B.); The ALS association (D.W.C., M.W.B., and S.V.-S.); MDA development grants (Z.M. and J.L.-E.); The BrightFocus Foundation (A.R.A.d.A.Q.); and Cancer Center Support Grant CA034196 to The Jackson Laboratory.


Loss of nuclear TDP-43 is a hallmark of neurodegeneration in TDP-43 proteinopathies, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). TDP-43 mislocalization results in cryptic splicing and polyadenylation of pre–messenger RNAs (pre-mRNAs) encoding stathmin-2 (also known as SCG10), a protein that is required for axonal regeneration. We found that TDP-43 binding to a GU-rich region sterically blocked recognition of the cryptic 3′ splice site in STMN2 pre-mRNA. Targeting dCasRx or antisense oligonucleotides (ASOs) suppressed cryptic splicing, which restored axonal regeneration and stathmin-2–dependent lysosome trafficking in TDP-43–deficient human motor neurons. In mice that were gene-edited to contain human STMN2 cryptic splice-polyadenylation sequences, ASO injection into cerebral spinal fluid successfully corrected Stmn2 pre-mRNA misprocessing and restored stathmin-2 expression levels independently of TDP-43 binding.