"GENETIC BASIS OF RARE DISEASES: ADVANCES IN DIAGNOSIS AND THERAPEUTIC APPROACHES"

Abstract

Mutation disabilities are behind rare diseases that cause heavy barriers to diagnosis and therapy. The study deals with rare disease genetics through diverse molecular techniques, which include mutation screening based on PCR, NGS, CMA, gene expression profiling, and CRISPR-Cas9 Gene editing, etc. Genomic DNA of good quality was extracted from the patient's samples and targeted sequencing has identified multiple pathogenic mutations. Whole-exome sequencing (WES) in combination with CMA has brought about the identification of some novel single nucleotide polymorphisms (SNPs), copy number variations (CNVs), as well as structural rearrangements, some of which are without previous reports. Analysis of gene expression has shown considerable dysregulation across major genes associated with the disease, with effective validation by Western blot matching alteration with protein levels. Functional validation by CRISPR-Cas9 was successful in restoring both normal gene and protein functions by correcting pathogenic mutations, careful statistical analysis presents a strong association of genetic alterations with disease phenotypes that confirm the clinical importance of these mutations identified. This study stresses the blend of multiple techniques for molecular diagnosis and treatment advancement even though variants may still be labeled uncertain. Findings pave the way for targeted gene therapies for rare diseases, thereby advancing the field of precision medicine. Future efforts should concentrate on widening patient cohorts and new treatment paradigms to improve treatment outcomes for individuals suffering from genetic diseases. This study stresses the knowledge of genetic pathophysiology and its application to personalized medicine.