Rational Design of ssODN to Correct Mutations by Gene Editing

Gene editing allows to make a variety of targeted changes in genome, which can potentially be used to treat hereditary human diseases. Despite numerous studies in this area, effectiveness of gene editing methods for correcting mutations is still low, so these methods are not allowed in routine practice. It has been shown that rational design of genome editing components can significantly increase efficiency of mutation correction. In this work, we propose design of single-stranded oligodeoxyribonucleotides (ssODNs) to increase efficiency of gene editing. Using a model system to repair knocked out EGFP that is integrated into the genome of HEK293T cell culture, we have shown that only a small part of ssODN (about 20 nucleotides: from the 15th nucleotide at 3 '-end to the 4th nucleotide at 5 '-end), a donor molecule for repairing double-stranded DNA breaks, is integrated into the site of the break. Based on the obtained data, it is possible to rationally approach the design of ssODNs to correct mutations using CRISPR-Cas9 method for the development of gene therapy for hereditary human diseases.

Automated summary

Gene editing allows targeted changes in the genome, potentially treating hereditary diseases. However, the effectiveness of gene editing methods in correcting mutations is still low. This study proposes the design of single-stranded oligodeoxyribonucleotides to improve the efficiency of gene editing, providing insights for developing gene therapy for hereditary diseases.