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Control Of Protease Activity In S. Pombe Using Gene Silencing Approach | 38967
Journal of Biotechnology & Biomaterials
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Amongst yeasts, the fission yeast Schizosaccharomyces pombe, is an attractive host model for high-level protein production
and functional analysis of eukaryotic proteins as it shares many molecular, genetic and biochemical features with higher
eukaryotes such as plants and animals, and is distinguishable from other yeasts through its ability to proliferate by fission rather
than budding. Furthermore, S. pombe has a developed Golgi apparatus and galactosyl transferase that is not found in other
yeast cells. However, one of the major hurdles in efficient production and purification of heterologous proteins from S. pombe is
proteolytic degradation of the recombinant gene products by host-specific proteases. The problem becomes significant when the
recombinant protein under production, is secretory and proteolytically sensitive in nature. The present study aims at controlling
the protease activity by gene silencing approach. RNA silencing is an evolutionarily conserved gene regulatory mechanism
with many species-specific variations by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA
sequences. It has become a powerful tool for genetic analyses and is likely to become a potent therapeutic approach for gene
silencing. A Protease silencing cassette was designed to impede the protease enzyme post trascriptionally. Since all proteases
do not attack all proteins, only protein specific protease is sought to be silenced as a test case in this study. Human parathyroid
hormone having a chain of 115 AAs was selected as model protein and the silencing of protease responsible for its degradation
was studied as a test case.
Nipunjot Kaur has been working as an Assistant Professor in the Department of Biotechnology Khalsa College, Punjab, India. She obtained her PhD from Department of Biotechnology Punjabi University in the year 2014.