Abstract:
Cystic Fibrosis, an autosomal recessive disorder of genetic origin, is primarily caused by mutations in the cystic fibrosis transmembrane conductance regulator gene. The mutated gene produces CFTR protein that is unable to maintain ion balance across cell membranes. More than 2,000 mutations have been identified in the CFTR gene. The cystic fibrosis transmembrane conductance regulator gene (CFTR gene), located at chromosome number 7, encodes for CFTR protein. The malfunctioning of CFTR protein causes irregularities in cellular electrolytes and water balance that leads to thickness of secretions and ultimately clog the airways and glandular ducts. Severity of this condition leads to death of individuals by affecting multiple organs especially lungs and pancreas. The mutated gene produces CFTR protein that is unable to maintain ion balance across cell membranes. More than 2,000 mutations have been identified in the CFTR gene. However, Pakistan's mutational spectrum is still unknown. The prevalence of Cystic Fibrosis (CF) in Pakistan is not known to certainty. CF is under diagnosed and under reported disease in Pakistan due to many reasons. Cystic Fibrosis disease causative mutations are rarely studied in Pakistan, especially no studies available on local populations of Southern Punjab Pakistan. Therefore, current study was persuaded to sequence the exon 11 of CFTR gene in CF patients of Southern Punjab region, Pakistan. The exon 11 of CFTR gene hosts ?F508 mutation which is most prevalent mutation of CFTR gene and exist in more than 66% CF patients of whole world and even up to 100% CF patients of some of isolated/conserved populations as of Fore Islands (Denmark). To fulfill the objectives of this study, Sanger sequencing technique was applied to sequence the exon 11 to detect ?F508 CFTR gene mutation or if any other mutation on exon 11 in local population of Southern Punjab, Pakistan. Genomic DNA of 12 CF patients was extracted from blood samples. Two primers, each of 21 bp designed through Primer-3 software and used to PCR amplify the exon 11 of CFTR gene of all DNA samples in 306bp PCR amplicons through thermal cycler (PCR machine). PCR products (306bp) of exon 11 was sequenced through Sanger sequencing/capillary electrophoresis. The sequenced results were analyzed for mutational investigation which predicted that ?F508 mutation or any other causative mutation in all 12 Cystic Fibrosis patients. That one is homozygous ?F508 out of 12 patients. These results suggest that ?F508 mutation of exon 11 is uncommon in population under study unlike other global populations due to genetic diversity in different ethnic populations. Availability of samples for recent research work was a challenging task as CF is a rare disease, so, a large-scale genomic study is recommended to find the mutational spectrum of whole CFTR gene and develop an effective mutational profile for better disease management and its treatment in Pakistan.
Page(s):
223-225
DOI:
DOI not available
Published:
Journal: 4th International Conference of Sciences “Revamped Scientific Outlook of 21st Century, 2025” , November 12,2025, Volume: 1, Issue: 1, Year: 2025
Keywords:
Pakistan
,
Southern Punjab
,
Cystic fibrosis
,
Sanger Sequencing
,
F508 mutation
,
CFTR Gene