Date of Award

12-7-2022

Degree Type

Thesis

Degree Name

MPhil in Biological and Biomedical Sciences

First Advisor

Dr. Ambrin Fatima

Second Advisor

Dr. Salman Kirmani

Third Advisor

Miss Fizza Akbar

Department

Biological and Biomedical Sciences

Abstract

Nearly 3.5-5.9% of the global population is affected by rare diseases at any point in time, which equates to 263-446 million affected people worldwide. Approximately 80% of all rare diseases have genetic origin with many having undetermined genetic etiology and mechanism which limits diagnosis and treatment. This project aimed to investigate the underlying cause of rare disorders exhibiting Mendelian pattern of inheritance in Pakistani population by using cutting edge sequencing techniques and molecular tools that would directly benefit the affected families. Two different diseases i.e. Severe Combined Immunodeficiency (SCID) and Developmental Disability had been identified in two different consanguineous families. Both diseases were found to segregate in respective family with Autosomal Recessive mode of inheritance. In first family (Chapter II) novel, missense, homozygous variant of the gene DCLRE1C c.767G>A (p.Cys256Tyr) was identified using Gene Panel Sequencing (GPS). Mutations in DCLRE1C gene have been previously linked with Severe Combined Immunodeficiency (SCID). Therefore, to determine the pathogenicity of the identified variant, Insilico tools were used to predict the effect of variant. With the aim to further investigate the pathophysiological mechanism associated with the variant, wild type (G/G) and mutant (A/A) DCLRE1C expression constructs (Site Directed Mutagenesis) were developed using Restriction Cloning. The developed constructs will be used for functional studies in future to validate the effect of the variant. In second family (Chapter III) two novel, missense, homozygous variants of the genes; CCDC47 c.1229C>T (p. Arg410His) and RTN4R c.1195G>A (p. Arg399Try) were identified using Whole Exome Sequencing (WES) due to diffuse phenotype showing Developmental Disability. Both the variants were found to segregate with the disease phenotype. Further functional studies will help to identify and validate contribution of these variants to disease phenotype. These studies have expanded the mutation spectrum of both rare Mendelian disorders in Pakistani population, having potential to translate into development of new diagnostic and treatment strategies to improve patient care in future.

First Page

1

Last Page

87

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