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Regulatory Challenges for Gene Delivery
Published in Yashwant Pathak, Gene Delivery, 2022
Vineet Mahajan, Shruti Saptarshi, Yashwant Pathak
Plant genome editing has the potential to offer advantageous traits to global agriculture. The CRISPR-Cas system is the most preferred gene editing approach that enables precise genome manipulation to achieve improved plant growth, nutrition, disease control, or sustainability. Nevertheless, there are inconsistencies in the regulatory framework covering gene-edited crops world-wide, thus impacting the public acceptance and marketability of these new plant varieties. Extension of currently established regulations covering conventional GMOs to the products generated through gene editing is a matter of debate needing further elucidation.
Nucleic Acids as Therapeutic Targets and Agents
Published in David E. Thurston, Ilona Pysz, Chemistry and Pharmacology of Anticancer Drugs, 2021
Although the CRISPR-Cas system is comparatively efficient and accurate, like other gene-editing approaches it may still cause off-target effects (e.g., cutting other, non-targeted, sequences of DNA), leading to concerns about the possibility of unwanted genomic damage that could cause cancers unrelated to the one being treated. In addition, there are ethical concerns that CRISPR-Cas-9 will be used to make germ line cell modifications which could permanently change genetic information in future generations. This fear was realized in 2018 when a Chinese researcher, He Jiankui, an associate professor at the Southern University of Science and Technology in Shenzhen (China) shocked the world by announcing that he had used CRISPR-Cas9 technology to modify the DNA of human embryos for HIV resistance, and that this had led to the birth of twin girls, Lulu and Nana. In 2019, He Jiankui was jailed for three years.
Balancing social justice and risk management in the governance of gene drive technology
Published in Christine Hauskeller, Arne Manzeschke, Anja Pichl, The Matrix of Stem Cell Research, 2019
Together, these factors are likely to increase shared global risks of gene drive applications, and to constitute a significant challenge to the consistent control of this technology field. While the above-mentioned scenarios are at present merely hypothetical, some of these developments can easily become reality in the near future. For policy-makers and scientists, it will be vital to take these possibilities into account, to contemplate the risks of these possible developments, and to integrate these insights into international debates and policies. One of the key challenges for the consistent governance of gene drive (and other gene editing) technology is that with the arrival of the CRISPR-Cas system, genetic modifications of living organisms are relatively easy to perform and at a low cost. As Jennifer Doudna, one of the inventors of the CRISPR technology, has remarked, in principle a graduate student with access to basic laboratory equipment could engineer a GM organism and release it into the environment (Baltimore et al., 2015). This could theoretically induce gene drives in biological populations, entirely without external controls, approval, or monitoring.
Shaping the future from the small scale: dry powder inhalation of CRISPR-Cas9 lipid nanoparticles for the treatment of lung diseases
Published in Expert Opinion on Drug Delivery, 2023
Simone P. Carneiro, Antonietta Greco, Enrica Chiesa, Ida Genta, Olivia M. Merkel
One of the main determinants of the safe use of the CRISPR-Cas system is the ‘off-target mutation’ referring to unsolicited and uncontrolled changes resulting in genome instability, loss of efficacy, and activation of pathological pathways; off-target genome modifications detected in human and mammalian systems cannot be overlooked. One of the strategies adopted to address this condition was studied by Slaymaker [32] where the Cas protein was skillfully modified to enhance the ability of the Cas domains to specifically recognize and interact exclusively with the targeted DNA. Another approach is represented by the combination of laboratory techniques and Machine learning [33]. Finally, CRISPR-Cas delivery strategies are widely studied to limit the drawback of off-target effects [34].
An overview: CRISPR/Cas-based gene editing for viral vaccine development
Published in Expert Review of Vaccines, 2022
Santosh Bhujbal, Rushikesh Bhujbal, Prabhanjan Giram
One of the most widely reported limitations of the CRISPR/Cas system is the off-target effects. In the interaction of sgRNA and target DNA, the RNA-guided nucleases employed in CRISPR methods were found to interact with numerous mismatch patterns [168]. These off-target effects possess the ability to cause genetic mutations or chromosomal rearrangements that aren’t intended. Many parameters impacting off-target CRISPR\Cas9 editing have been uncovered in a number of recent investigations to reduce off-target mutations. The primary parameters impacting off-target effects include sequence complementarity, PAM recognizing selectivity, target gene similarities, and Cas9 expression rate. Other factors, including the fidelity of gRNA attachment and the availability of the target, may also impact the chance of off-target altering [169,170]. As a result, a variety of tactics were used to figure out how to improve specificity and decrease off-target effects [171,172]. When it gets to viral genetic engineering, even so, off-target effects are less likely. Numerous viral editing findings show no potential off-target effects because the genome of viruses is much lesser than the genome of the host. This is worth noting, even so, that multiple genes may be cleaved as well as repaired in genomic sequences with overlapping genes [173].
Diagnostic accuracy of CRISPR technology for detecting SARS-CoV-2: a systematic review and meta-analysis
Published in Expert Review of Molecular Diagnostics, 2022
Xin Li, Huiling Zhang, Jing Zhang, Yang Song, Xuening Shi, Chao Zhao, Juan Wang
In 2013, Zhang et al. have firstly reported CRISPR-Cas Systems used in multiplex genome engineering, and later CRISPR technology came to apply for developing detection technology [8,9]. Diagnostic technology based on CRISPR-Cas can simultaneously satisfy a variety of detection criteria, which has the potential to be used to the next generation of diagnostic technology [10,11]. CRISPR Cas system can edit target DNA or RNA sequences with CRISPR Cas enzymology and amplification process in disease diagnosis platforms. Firstly, the vast amount of target nucleic acids synthesized with an amplification process, such as RT-RPA (reverse transcription recombinase polymerase amplification) or RT-LAMP [12]. Secondly, CRISPR RNA (crRNA) identifies the target nucleic acid for the spacer of the CRISPR Cas system and then the target nucleic acid was cut by specific Cas nucleases including Cas 9, Cas12 and Cas13 nucleases [12–14]. Finally, target nucleotides testing result was output by signal readout, such as fluorescence detection, lateral flow assay (LFA), and colorimetric aggregation with nanoparticles [12,15,16]. A large number of studies have reported that CRISPR technology has the advantages of low cost, ease to use, high sensitivity and specificity. To a certain extent, it can overcome the limitations of traditional molecular diagnostic methods such as RT-qPCR [17–19].