## 4 Discussion  
Patients suffering from epidermolysis bullosa (EB) face lifelong discomfort and treatment dilemmas, particularly in Pakistan, where health care facilities have poor availability. Rare genetic diseases are more frequent in this population because of a deficit in genetic testing facilities and poor information for diseased families. All types of DEB are caused by genetic defects of the \(COLYA1\) gene, which translates type VII collagen [12]. Type VII collagen is coded by \(COLYA1\), spanning 118 exons on 3p21.10 chromosome. The mRNA transcript of \(\sim\)8.9 kb produce procollagen \(\alpha\)1 (VII) chain consisting of 2944 amino acids [12]. The three main structures of each polypeptide chain are: a collagenous triple helix (coded by 29 to 112 exons), an amino terminus NC-1 (coded by exons 2 to 28 exons) and a carboxyl terminus NC-2 (coded by 113 to 118 exons) [13]. The formation of anchoring fibrils takes place by deposition of collagen VII underneath the basement membrane of the epidermis of the skin where it is responsible for the connection of dermal ECM to the basement membrane. Deficient collagen VII causes persistent fragility of the skin, ongoing fibrosis of multiple organ systems, and scarring [14]. In severe generalized RDEB, homozygous or compound heterozygous mutations in the \(COLYA1\) gene produce premature termination codons, leading to absence of anchoring fibril production due to degradation of abnormally truncated polypeptides [15]. [PERSON] [16] created a transgenic mouse model with contingent inactivation of \(COI7A1\) expression, resulting in a \(COI7A1\) hypomorphic animal expressing about 10% of normal \(COI7A1\) and clinical symptoms that closely resemble human recessive abnormalities, such as nail dystrophy, skin fragility, and growth failure. Clinically, RDEB-GS patients present with the involvement of the integumentary system, including the mucous membranes (Figure 2), \begin{table}
\begin{tabular}{c c c c c c c c c} \hline
**Gene** & **Variant** & **MT** & **PolyPhen2** & **MUPRO** & **Fathmm** & **VarsSome** & **PROVEAN** & **MutPred** & **SIFT** \\ \hline COI7A1 & c,2034C\(\alpha\)A & Disease & Possibly & Decrease & Damping & Likely & Deuterous & Pathogenic & Damping \\  & p,C3p34C\(\alpha\)A & causing & Damping & stability & Damping & Dalephonic & Datterous & Pathogenic & Damping \\ COI7A1 & p,Hil2034 Raffect178 & causing & NA & NA & Pathogen & NA & NA & NA \\ COI7A1 & p,K52 NC\(\alpha\)T & Dosen & NA & NA & Neutral & Pathogenic & NA & NA & NA \\ \hline \end{tabular}
\end{table} 
Table 2: Pathogenicity detection of \(COLYA1\) variants. Figure 4: Multiple sequence alignment of Human Gly2345 in Family **A**, Human His129 in Family **B** and Human Arg525 in Family **C** with their orthologs. Red arrows showing their evolutionary conservation through different species. esophageal structures necessitating gastrostomy tube placement, mitten deformities of the hands, toes and fingers, loss of nail plates, joint contractures, and eye inflammation with visual impairment [17]. Additionally, these excessive heavily fibrotic lesions favor the progressive formation of aggressive cutaneous squamous cell carcinoma (SCC) at an early age, the peritumoral infiltration of immune cells causes immunosuppression and metastasis development [18; 19], decreasing their life expectancy to only their fourth decade [20]. Many of the same clinical characteristics are present in RDEB-GO individuals; however, they are milder. The non-severe variants have a little better prognosis, with a median survival age of 55 to 65 years, basically because of immunosuppression and immunosuppressive which are responsible for onset and progression of malignant lesions at the molecular level [21]. Although a cure for EB is still a long way off, recent findings in clinical trials based on humans and animal models have encouraged affected individuals, clinicians, and genetic investigators to believe that modification of the disease factors and enhancement of quality of life are achievable motives [4; 14].