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Hatami bardar A, Azimi nejad M, Jafarinia M, Goodarzi H R, Mojarrad M. Generation of IDUA knockout cell lines by CRISPR/Cas mediated genome editing. yafte 2021; 22 (4) :162-169
URL: http://yafte.lums.ac.ir/article-1-3103-en.html

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.