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Genome Editing Tools
Published in Vineet Kumar, Vinod Kumar Garg, Sunil Kumar, Jayanta Kumar Biswas, Omics for Environmental Engineering and Microbiology Systems, 2023
Madhumita Barooah, Dibya Jyoti Hazarika
Many industrially important products have been produced by overexpressing heterogeneous pathways in different bacteria. The successful application of CRISPR-Cas technology for metabolite production includes n-butanol and 5-aminolevulinic acid production in E. coli (Ding et al., 2017; Heo et al., 2017), n-butanol production in Clostridium saccharoperbutylacetonicum (Jiménez-Bonilla et al., 2021), isopropanol-butanol-ethanol production in Clostridium acetobutylicum and C. saccharoperbutylacetonicum (Wasels et al., 2017; Wang et al., 2019), succinic acid production in Synechococcus elongatus (Li et al., 2016), and γ-aminobutyric acid (GABA) production in Corynebacterium glutamicum (Cho et al., 2017). In Corynebacterium glutamicum, Cas12a (formerly Cpf1)-based RNA-guided endonuclease was also used to enhance the natural proline production through site-directed mutagenesis to dismiss the product inhibition (Jiang et al., 2017). Success stories of CRISPR-Cas platform for bacterial metabolic engineering may expand its applicability in the remediation of natural pollutants under controlled environment.
Genetic Engineering as a Tool for Enhanced PHA Biosynthesis from Inexpensive Substrates
Published in Martin Koller, The Handbook of Polyhydroxyalkanoates, 2020
Lorenzo Favaro, Tiziano Cazzorla, Marina Basaglia, Sergio Casella
Recently, the development of the proficient gene-editing CRISPR/Cas9 technology paved the way for novel and extremely interesting research tools. The genome-editing methods based on CRISPR/Cas9, CRISPR-Cas12a, and/or CRISPRi have been recently reviewed as becoming increasingly crucial for the regulation of metabolic flux to PHA, the development of strong PHA synthetic pathways, and further host strain optimization [133]. The simultaneous integration of genes into multiple loci of the host genome via CRISPR/Cas9 will support researchers in the near future to edit microbial genomes more quickly, targeting both several PHA syntheses and multiple substrate-utilization pathways to be promptly improved.
Crispr biosensing and Ai driven tools for detection and prediction of Covid-19
Published in Journal of Experimental & Theoretical Artificial Intelligence, 2023
Abdullahi Umar Ibrahim, Pwadubashiyi Coston Pwavodi, Mehmet Ozsoz, Fadi Al-Turjman, Tirah Galaya, Joy Johnson Agbo
A study reported by Pardee et al. (2016) integrated CRISPR/Cas9 with optical geno-biosensor to develop a CRISPR-paper-based biosensor for detection of Zika virus at fM concentration. The sensitivity of the biosensor is as a result of hybridisation of Zika virus RNA with gRNA. The research employed Dengue virus as a negative control, and the result has shown the precision of the biosensor to differentiate between Zika virus and dengue virus and viral strains based on SBP. Due to the high need of cheap and portable biosensor, Dai et al. (2019) utilised the trans-cleavage activity of Cas12a enzyme derived from Lachnospiraceae bacterium ND2006 Cas12a (LbCas12a) and Acidaminococcus sp. (AsCas12a) to develop an electrochemical CRISPR-based biosensor for detection of HPV-16 and parvovirus B19. The biosensor archived 50pM sensitivity LOD. J. S. Chen et al. (2018) harnessed the RNA-guided DNA-binding activity of CRISPR/Cas12a (Cpf1) derived from Lachnospiraceae bacterium ND2006 Cas12a (LbCas12a) combined with isothermal amplification method to amplify target DNA to develop a DNA endonuclease-targeted CRISPR, transreporter (DETECTR), which is a molecular diagnostic method for the detection of HPV16 and HPV18 from patient samples. The system achieved attomolar sensitivity for detection of target NA.