Fungal genome editing using g CRISPR-Cas nucleases: a study for enhanced cellulases production | [Abstract Book on International Conference on Food and Applied Sciences (ICFAS-23) 3-5 August 23 • 2023]

Author(s):

1. Riffat Seemab: Department of Bioinformatics and Biotechnology, Government College University Faisalabad (GCUF), Allama Iqbal Road, Faisalabad-38000, Pakistan

2. Zeeshan Shokat: Department of Bioinformatics and Biotechnology, Government College University Faisalabad (GCUF), Allama Iqbal Road, Faisalabad-38000, Pakistan

3. Abdul Zahir Abbasi: Department of Bioinformatics and Biotechnology, Government College University Faisalabad (GCUF), Allama Iqbal Road, Faisalabad-38000, Pakistan; Department of Biotechnology, University of Azad Jammu and Kashmir, Chehla Campus, 13100-Muzaffarabad-AJK, Pakistan

4. Muhammad Rizwan Javed: Department of Bioinformatics and Biotechnology, Government College University Faisalabad (GCUF), Allama Iqbal Road, Faisalabad-38000, Pakistan

5. Anam Ijaz: Department of Bioinformatics and Biotechnology, overnment College University Faisalabad (GCUF), Allama Iqbal Road, Faisalabad, Pakistan

Abstract:

Aspergillus nigerhas beenexplored as an industrially important fungus that is Generally Recognized as Safe (GRAS). Presence of specific and content enriched carbohydrate-active enzyme families that are involved in plant biomass degradation, makes this strain a promising industrial cell factory for cellulases enzyme production. Cellulases (exo-glucanases, endo-glucanases, and ß-glucosidases) are among the most in-demand commercial industrial enzymes due to their wide range of potential uses in industries like pulp and paper, textiles, food processing, agriculture, biotechnology. Several transcriptional factors (activators & repressors) are known to work synergistically for cellulases expression in fungi. Over the pastdecade, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 (CRISPR-associated nuclease 9) has emerged as a ground-breaking technology in genome engineering due to its high efficiency and versatility. In the envisaged study to achieveenhanced cellulase enzymes titre, CRISPR-Cas9 based recombinant plasmid was constructed harbouring an AMA1-based autonomously replicating vector, hygromycin (hph) resistance marker and gRNAto introduce mutations/silencing of the repressor geneto inhibit catabolite repression. Protoplast-based transformation system was developed for genome editing. The engineered (?repressor gene) strain exhibited two-fold improvement in cellulases production as compared to the wild strain. The optimized method will further enable todevelop novel strains to produce heterologous enzymes and improvement of their metabolic pathways.

Page(s): 103-103

DOI: DOI not available

Published: Journal: Abstract Book on International Conference on Food and Applied Sciences (ICFAS-23) 3-5 August 23, Volume: 0, Issue: 0, Year: 2023

Keywords:

Aspergillus niger , CRISPRCas9 , Repressor gene , Cellulases , GRAS

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