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Treatment of a Mouse Model of ALS by In?Vivo Base Editing

Mol Ther. 2020; 
Lim CKW, Gapinske M, Brooks AK, Woods WS, Powell JE, Zeballos C MA, Winter J, Perez-Pinera P, Gaj T.
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Recombinant Proteins 23 The following primary antibodies were used: rabbit anti-hSOD1 (1:200; Cell Signaling Technology, 2770S), goat anti-ChAT (1:25; EMD Millipore; AB144P), goat anti-HA (1:250; GenScript, A00168), rabbit anti-HA (1:500; Cell Signaling Technology, 3724S), chicken anti- HA (1:500; Abcam, ab9111), rabbit anti-FLAG (1:500; Cell Signaling Technology, 14793S) rabbit anti-NeuN (1:500; Abcam, ab177487), rabbit anti-IbaI (1:500; Wako Chemicals, 019- 19741), rat anti-Mac2 (1:500; Cedarlane Labs, CL8942AP), mouse anti– β3-tubulin (1:1000; Sigma-Aldrich, T8578), chicken anti-GFAP (1:1000; Abcam, ab4674) and rabbit anti- synaptophysin 1 (1:500; Synaptic Systems, 101 002). Get A Quote

摘要

Amyotrophic lateral sclerosis (ALS) is a debilitating and fatal disorder that can be caused by mutations in the superoxide dismutase 1 (SOD1) gene. Although ALS is currently incurable, CRISPR base editors hold the potential to treat the disease through their ability to create nonsense mutations that can?permanently disable the expression of the mutant SOD1 gene. However, the restrictive carrying capacity of adeno-associated virus (AAV) vectors has limited their therapeutic application. In this study, we establish an intein-mediated trans-splicing system that enables in?vivo delivery of cytidine base editors (CBEs) consisting of the widely used Cas9 protein from Streptococcus pyogenes. We show that intrathecal... More

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