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

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.

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.

Analytical digestion and Sanger sequencing confirmed indel mutation induction in two clones in heterozygote and one clone in homozygote state.

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.