Towards Optimizing Acetic Acid Utilization In Saccharomyces cerevisiae Through gRNA-Landing Pads
Typ
Examensarbete för masterexamen
Master's Thesis
Master's Thesis
Program
Biotechnology (MPBIO), MSc
Publicerad
2024
Författare
Berg, Erik
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
Lignocellulosic biomass, residual biomass from the agricultural and forestry industries, is the most
abundant organic substance on the planet. For efficient utilization by Saccharomyces cerevisiae, the
biomass must undergo a pretreatment process that typically generates strong inhibitors, with acetate
being the most prominent. This project served as a proof of concept and explored optimization of
acetate utilization in Saccharomyces cerevisiae using a gRNA-landing pad strategy. This was done
through a wide range of molecular biology methods such as the transformation of Escherichia coli and
the Saccharomyces cerevisiae strain LX6, restriction cloning, cloning using the MoClo system, as well
as in-silico design of plasmids and primers. A gRNA-landing pad was successfully assembled in-vitro,
using in-silico custom-made MoClo parts, and transformed into Escherichia coli. Transformation of
the gRNA-landing pad into LX6 presented a challenge, that as of yet is not overcome. Despite the
challenges in transforming the yeast, the creation of an easy-to-assemble gRNA-landing pad could
enable a fast and reliable method for future research involving gRNA-landing pads Saccharomyces
cerevisiae.