Step-wise approach for In silico CRISPR-Cas9 gene editing of IVS I-110 and IVS I-6 Beta-thalassemia-causing variants

Amr, Khalda Said; Rabie, Eman Abdelalim; Dawood, Reham; El-Kamah, Ghada

doi:10.21608/ejchem.2024.304964.10042

Step-wise approach for In silico CRISPR-Cas9 gene editing of IVS I-110 and IVS I-6 Beta-thalassemia-causing variants

Document Type : Original Article

Authors

¹ Medical Molecular Genetics Department, National Research Centre, El-Buhouth St., Dokki, 12622 Cairo, Egypt

² Medical Molecular Genetics Department, Human Genetics & Genome Research Division (HGGR), National Research Centre (NRC), Cairo 12622

³ Microbial Biotechnology Department, National Research Centre, Dokki, Cairo, Egypt

⁴ Clinical Genetics Department, Human Genetics & Genome Research Division (HGGR), National Research Centre (NRC), Cairo 12622, Egypt

10.21608/ejchem.2024.304964.10042

Abstract

β-thalassemia is a common monogenic blood disorder caused by disease-causing variants, mostly point mutations in the HBB gene. In Egypt, it is the most common hemoglobinopathy with a minor allele frequency of 0.03 and a carrier rate varying between 5.3 to more than 9% creating a major social and financial burden for the patients, their family, and the society in general. The current study aims to design a guide RNA to target the most common mutations identified among B Thalassemia patients. By using the human variation viewer database, two mutations within the HBB gene have been selected. By computational tools, two guide RNA have been designed and showed a high-efficiency score towards the desired mutations. The selected two gRNA were successfully cloned into a synthetic plasmid(PX459).

The data successfully established a step-wise in-silico approach for designing a CRISPR/Cas9 gene editing model for the correction of IVS I-110 (G>A, rs35004220) and IVS I-6 (T>C, rs35724775) variants in HBB gene.

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