In Vivo Repair of a Protein Underlying a Neurological Disorder by Programmable RNA Editing
John R. Sinnamon,
Susan Y. Kim,
Jenna R. Fisk,
Zhen Song,
Hiroyuki Nakai,
Sophia Jeng,
Shannon K. McWeeney,
Gail Mandel
Affiliations
John R. Sinnamon
Vollum Institute, Oregon Health and Science University, Portland, OR 97239, USA
Susan Y. Kim
Vollum Institute, Oregon Health and Science University, Portland, OR 97239, USA
Jenna R. Fisk
Vollum Institute, Oregon Health and Science University, Portland, OR 97239, USA
Zhen Song
Department of Molecular and Medical Genetics, Oregon Health and Science University, Portland, OR 97239, USA
Hiroyuki Nakai
Department of Molecular and Medical Genetics, Oregon Health and Science University, Portland, OR 97239, USA
Sophia Jeng
Division of Bioinformatics and Computational Biology, Department of Medical Informatics and Clinical Epidemiology, Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA
Shannon K. McWeeney
Division of Bioinformatics and Computational Biology, Department of Medical Informatics and Clinical Epidemiology, Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA
Gail Mandel
Vollum Institute, Oregon Health and Science University, Portland, OR 97239, USA; Corresponding author
Summary: Programmable RNA editing is gaining momentum as an approach to repair mutations, but its efficiency in repairing endogenous mutant RNA in complex tissue is unknown. Here we apply this approach to the brain and successfully repair a guanosine-to-adenosine mutation in methyl CpG binding protein 2 RNA that causes the neurodevelopmental disease Rett syndrome. Repair is mediated by hippocampal injections of juvenile Mecp2317G>A mice with an adeno-associated virus expressing the hyperactive catalytic domain of adenosine deaminase acting on RNA 2 and Mecp2 guide. After 1 month, 50% of Mecp2 RNA is recoded in three different hippocampal neuronal populations. MeCP2 protein localization to heterochromatin is restored in neurons to 50% of wild-type levels. Whole-transcriptome RNA analysis of one neuronal population indicates that the majority of off-target editing sites exhibit rates of 30% or less. This study demonstrates that programmable RNA editing can be utilized to repair mutations in mouse models of neurological disease.
