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Microalgae as a Source of Sustainability
Published in Pau Loke Show, Wai Siong Chai, Tau Chuan Ling, Microalgae for Environmental Biotechnology, 2023
Pik Han Chong, Jian Hong Tan, Joshua Troop
Besides improving the infrastructure, advancement on microalgae is also achievable. Genetic engineering of microalgae can help overcome or bypass the biological limitations of its metabolic capacity. Genetic traits such as higher accumulation of desired biomolecules, improved photosynthetic productivity, respective cellular production, production of value-added compounds, or just purely increasing algal biomass productivity (Fu et al. 2019) aim to improve the economic feasibility of the production process. There are recent advances in novel gene-editing tools such as zinc-finger nuclease (ZFN), TAL effector endonuclease (TALEN), and clustered regularly interspaced palindromic sequences (CRISPR/Cas9). These gene-editing tools can facilitate highly specific targeting of genes for editing, allowing us to alter microalgae genome toward designed properties for various applications (Wang et al. 2016; Jeon et al. 2017).
Nanotechnology Applications in Nanomedicine: Prospects and Challenges
Published in Khalid Rehman Hakeem, Majid Kamli, Jamal S. M. Sabir, Hesham F. Alharby, Diverse Applications of Nanotechnology in the Biological Sciences, 2022
Arpita Dey, Smhrutisikha Biswal, Somaiah Sundarapandian
Since the discovery of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) in 2012, gene editing has gained extensive research momentum. The CRISPR-Cas9 (CRISPR-associated protein 9) is a recently developed gene-editing tool, a technology inspired by bacteria for new treatment of genetic diseases or disorders. CRISPR is composed of a scissor-like protein called Cas9, and a guide RNA molecule called sgRNA. The sgRNA guides the Cas9 protein to reach the target gene in the nucleus to edit the mistakes with the host cells’ repair system’s help. But to deliver the gene-clipping tool CRISPR-Cas9 in the cytosol and then to the nucleus across the cell membrane directly and effectively overcoming the cell’s defense system is a significant challenge (Mout et al., 2017). In a recent report, CRISPR/Cas9-ribonucleoprotein (Cas9-RNP)-based genome editing was able to specifically target gene and avoid integrational mutagenesis (Mout et al., 2017). The study found that Cas9–sgRNA complex coengineered with the cationic arginine gold nanoparticles (ArgNPs) showed high efficiency (∼90%) toward direct cytoplasmic and nuclear delivery besides approximately 30% gene editing efficiency. The Cas9 protein was also designed with an atomic sequence to release the inside nucleus by tweaking the Cas9 protein, and the delivery process was real-time monitored using advanced microscopy.
The Current State of Non-Viral Vector–Based mRNA Medicine Using Various Nanotechnology Applications
Published in Yashwant V. Pathak, Gene Delivery Systems, 2022
Kshama Patel, Preetam Dasika, Yashwant V. Pathak
CRISPR-Cas9 was inspired by a bacterium that used genome editing. In this system, the bacteria use parts of the DNA from foreign viruses to create CRISPR arrays, which are the pieces that the bacteria remember from the virus.21 Then, when the bacteria sense the virus again, they will produce RNA from the CRISPR arrays to attack the DNA from the virus. Lastly, the Cas9 part will come into play where the DNA of the virus is dissembled and destroyed.21 This process is very similar to the lab in which genome editing is done. In the lab, scientists will make a tiny piece of RNA, which can then attach to the DNA of the target genome and the Cas9 enzyme.21 This RNA is then able to identify the DNA of the genome, and Cas9 dissembles the DNA and allows for new genetic material to be added or existing genetic material to be removed.21 Furthermore, this allows the scientist to construct the DNA to their will, allowing the perfect DNA structure to be created.
The roadmap towards cure of chronic hepatitis B virus infection
Published in Journal of the Royal Society of New Zealand, 2022
In contrast, HBx binds the HBV minichromosome and modifies the epigenetic regulation of cccDNA function. HBV X-inhibitors may be a potential approach to silence cccDNA without off-target effects. Direct gene editing approaches include cccDNA degradation through IFN-α/Lymphotoxin-β receptor agonist (via APOBEC3A/B), or cccDNA cleavage via nucleases including Zinc-finger nucleases (ZFNs), transcription-activator like effector nucleases (TALENS) and CRISPR/Cas9 approach. In in vitro experiments, combining Cas9 with guide RNAs which target conserved regions within HBV cccDNA, achieves profound reductions in cccDNA and all HBV proteins (Wang et al. 2017). In addition to destroying cccDNA, CRISPR/Cas9 will also remove HBV integrins from the host genome, thereby reducing any long-term risk of hepatocarcinogenesis. The important challenges facing the successful use of gene editing approaches in patients include how to measure efficacy (no standardised cccDNA quantification assay), how to deliver the drug efficiently to the nucleus of every infected hepatocyte and finally, how to mitigate the long-term risks of chromosomal translocation in otherwise healthy young people (Table 1).
Gene doping: Present and future
Published in European Journal of Sport Science, 2020
Rebeca Araujo Cantelmo, Alessandra Pereira da Silva, Celso Teixeira Mendes-Junior, Daniel Junqueira Dorta
Some techniques have been helpful to leverage research in the field of gene editing, increasingly contributing to gene editing development and improvement. Among these techniques, zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and particularly the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR associated protein 9 (CRISPR-Cas9) are noteworthy. These tools are applied in genetic manipulation and have diverse functions including gene introduction or replacement, precise DNA insertion or deletions of various lengths, and production of knock-in and knockout animals and plants. However, all these genome-editing techniques require several processes to evaluate and to regularize their practice so that the underlying risks are known and their misuse can be avoided in the future (Fears & Meulen, 2017; Mashimo, 2014).
The potential for the use of gene drives for pest control in New Zealand: a perspective
Published in Journal of the Royal Society of New Zealand, 2018
Peter K. Dearden, Neil J. Gemmell, Ocean R. Mercier, Philip J. Lester, Maxwell J. Scott, Richard D. Newcomb, Thomas R. Buckley, Jeanne M. E. Jacobs, Stephen G. Goldson, David R. Penman
The advent of CRISPR/Cas9 targeting technologies (Hsu et al. 2014) has given new life to the gene drive idea. CRISPR/Cas9 makes use of a prokaryotic system which allows cells to cut invasive DNA that has been encountered previously (Horvath & Barrangou 2010). The system consists of a nuclease, Cas9, that can be targeted to any sequence in the genome using a small RNA sequence called a guide RNA (gRNA), providing that target sequence sits next to a 2–6 base pair PAM motif (Horvath & Barrangou 2010). The combination of the Cas9 molecule, which cuts DNA to form double-stranded breaks, and specific gRNA that guide Cas9 to a particular sequence, provides the technology to cut DNA at specific locations (Beumer & Carroll 2014; Bassett & Liu 2014). Using gRNAs targeting a specific sequence in a pest genome, a gene drive mechanism using CRISPR/Cas9 would act in the same way as for HEGs (Esvelt et al. 2014).