Mitigating symptoms of the Aicardi-Goutières Syndrome through gene editing by CRISPR-Cas9 Restricted; Files Only
Krishnakumar, Siddharth (Spring 2025)
Abstract
Aicardi-Goutières Syndrome (AGS) is a severe type of leukodystrophy that causes neurodegeneration and developmental regression in infants. It is a genetic disorder primarily caused by mutations in RNASEH2 subunit genes and the TREX gene. When a gene in the RNASEH2 subunit is mutated, R-loops accumulate, causing inflammation in the brain and symptoms characteristic of AGS. Through CRISPR-Cas9, we corrected the G529A mutation in the RNASE2B gene in patient-derived induced pluripotent stem cells (iPSCs). DNA-RNA Immunoprecipitation Sequencing was used to quantify genome-wide R-loop accumulation, while RNA-Seq was used to identify transcriptional regulation. Our findings show that mutant cells exhibited significantly higher accumulation of R-loops compared to Healthy Control controls, particularly near genes involving neural development and the adaptive immune response. Gene editing through CRISPR-Cas9 reduced R-loop accumulation in 89.36% of genomic regions where R-loops were significantly increased in mutant cell lines. These findings indicate that R-loop accumulation could be the causal mechanism for AGS onset and progression and that CRISPR-Cas9 may serve as a valid therapeutic strategy to mitigate symptoms of AGS.
Table of Contents
Abstract 1
Background 6
Methods 11
Results 17
Discussion 30
References 36
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File download under embargo until 22 May 2027 | 2025-04-14 00:25:08 -0400 | File download under embargo until 22 May 2027 |
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