MB 555 Microbial Biotechnology

Cystic fibrosis is a defect caused by mutations that alter the functioning of the transporter in the cystic fibrosis transmembrane conductance regulator of epithelial cells, which changes the normal functioning of several organs (Magalhaes et al., 2017). In the late 1900s, when the disease was discovered, research was conducted to understand the underlying mechanism of the defect and develop ways to rectify the defect. 

Forward genetics involves identifying a phenotype that is defective and creating a model organism with these qualities to find ways that will correct the defect. Sequencing enabled the identification of various responsible mutations and enabled cloning (Taylor- Cousar et al., 2020). Cloning was a major step conducted to restore the CFTR functioning and brought about gene-based therapies. Later on, viral and non-viral approaches were adapted to deliver CFTR to apical cells and rectify the receptors (Magalhaes et al., 2017).

Viral models included the use of adenovirus, retroviral and lentiviral vectors—these models served by complementing CFTR in cystic fibrosis patients. Virus’s models were introduced to the ailing patient in various forms (Taylor- Cousar et al., 2020). For patients with respiratory challenges, they were introduced through breathing which ensured that the lungs were reached and the defective chloride channels corrected. MuLV- based vectors, an example of lentivirus, were used. It worked by integrating into dividing cells and stimulate the proliferation of epithelial cells to improve the ionic transfers (Magalhaes et al., 2017). These vector models were evaluated on their abilities to transduce airway cells in vivo and in vitro. Challenges faced involved finding the correct number of cells that needed to be rectified for the disease to be cured. Another challenge faced was an immunologic response towards the foreign viruses introduced in the body.

Reverse genetics is a technique used to determine what phenotype arises due to changes in a particular genetic sequence (Taylor- Cousar et al., 2020). Linkage analysis enabled the defective chromosome to be identified on chromosome seven. More than 100 various mutations have been identified as the molecular detection of cystic fibrosis. Among these genes is a deletion on 3basepair deletion mutation (Magalhaes et al., 2017). The CFTR amino acid sequence shows that it comes from a family of integral membrane transport proteins. This impairs the regular second messenger cAMP involved in chloride transport. This presents as concentrated secretions produced by epithelial cells of the lung, pancreas, sweat glands, and kidneys. Identification of these genes has led to advances in the medical field (Magalhaes et al., 2017).

Forward genetics aims to correct the defective gene by introducing viral and non-viral models to rectify the defective chloride channels (Magalhaes et al., 2017). Reverse genetics aims to map out the gene to find exactly where the mutation occurred. Substituted, circular, or single-stranded genes, both RNA and DNA base pairs, insert the correct template to the defective genetic fibro

References

Magalhães, M., Rivals, I., Claustres, M., Varilh, J., Thomasset, M., Bergougnoux, A., ... & De Sario, A. (2017). DNA methylation at modifier genes of lung disease severity is altered in cystic fibrosis. Clinical epigenetics, 9(1), 1-15.

Taylor‐Cousar, J. L., Evans, T. A., Cutting, G. R., & Sharma, N. (2020). Potentially lethal cystic fibrosis gene variant in the orangutan. American journal of primatology.

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