Generation of IDUA knockout cell lines by CRISPR/Cas mediated genome editing

Hatami bardar, ali and Aziminejad, mohsen and Jafarinia, mojtaba and Goodarzi, hamidreza and Mojarad*, majid (2021) Generation of IDUA knockout cell lines by CRISPR/Cas mediated genome editing. Yafteh, 22 (4).


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Background: The hurler syndrome is the most common form of lysosomal storage diseases (LSDs). In the present experiment, we intended to generate an IDUA targeting CRISPR-Cas system and deactivate the target gene using it. Materials and Methods: IUDA targeting sgRNA pair were cloned into the px335 plasmid and resulted plasmid identity was confirmed using restriction mapping. Recombinant plasmid was transfected into the human epithelial cell line, HEK-293. Mutated clones were screened by melting analysis and their mutation was characterized by Sanger sequencing. Results: Analytical digestion and Sanger sequencing confirmed indel mutation induction in two clones in heterozygote and one clone in homozygote state. Conclusion: In this study, we produced an IDUA knockout cell model using CRISPR-nCas9 system. This model can be used in the therapeutic approaches of this disease. Keywords: Hurler syndrome, CRISPR-Cas, Gene knock out, IDUA

Item Type: Article
Subjects: R Medicine > R Medicine (General)
Depositing User: mania jalilvand
Date Deposited: 11 May 2021 06:00
Last Modified: 11 May 2021 06:00

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