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.