CRISPR-Cas9-mediated gene editing in human MPS I fibroblasts.

Abstract

Mucopolysaccharidosis type I (MPS I) is a lysosomal storage disorder (LSD). It is caused by mutations in the IDUA gene, which lead to the accumulation of the glycosaminoglycans dermatan and heparan sulfate. The CRISPR-Cas9 system is a new and powerful tool that allows gene editing at precise points of the genome, resulting in gene correction through the introduction and genomic integration of a wildtype sequence. In this study, we used the CRISPR-Cas9 genome editing technology to correct in vitro the most common mutation causing MPS I. Human fibroblasts homozygous for p.Trp402* (legacy name W402X) were transfected and analyzed for up to one month after treatment. IDUA activity was significantly increased, lysosomal mass was decreased, and next generation sequencing confirmed that a percentage of cells carried the wildtype sequence. As a proof of concept, this study demonstrates that CRISPR-Cas9 genome editing may be used to correct causative mutations in MPS I. LIST OF ABBREVIATIONS.

• Authors: Talita Giacomet de Carvalho, Roselena Schuh, Gabriela Pasqualim, Felipe Matheus Pellenz, Eduardo Cremonese Filippi-Chiela, Roberto Giugliani, Guilherme Baldo, Ursula Matte
• Journal: Gene (Gene) Vol. 678 Pg. 33-37 (Dec 15 2018) ISSN: 1879-0038 [Electronic] Netherlands
• PMID: 30081189 (Publication Type: Journal Article)
• Copyright: Copyright © 2018 Elsevier B.V. All rights reserved.

Chemical References

• IDUA protein, human
• Iduronidase

Topics

• CRISPR-Cas Systems
• Cells, Cultured
• Fibroblasts (cytology, metabolism)
• Gene Editing (methods)
• Genetic Therapy
• High-Throughput Nucleotide Sequencing
• Humans
• Iduronidase (genetics)
• Mucopolysaccharidosis I (genetics, therapy)
• Mutation
• Sequence Analysis, DNA