Literature Review copy

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Human Genetic Engineering and the Potential Risk Factors Based on Ethics and Restrictions Many have discussed the advantages of human genetic engineering and how it can help future generations through the means of heath. However, many argue that human genetic engineering can have many disadvantages especially when the topic of ethics come up and the risk factors involved. Gene therapy or genetic engineering allows for the possibility for treatment of genetic diseases. One of the major discoveries of gene therapy was CRISPR/Cas 9 which paved the way for the development of base editing and future gene therapy discoveries (Godbout & Tremblay, 2023). The researches discussed addresses the issue of germline and somatic modifications (in addition to CRISPR and prime editing) and how they have presented risk factors such as, mutations and the lack of awareness when it comes to the potential consequences and results of gene editing. In 2019, there was a new discovery called prime editing. Prime editing could interfere with targeted deletion, insertions or all base-base conversions (Godbout & Tremblay, 2023). The primer binding sequence attached to the DNA flap and using the reverse transcriptase (RTT) the edited RNA sequence is reverse transcribed. Once this is complete, the edited strand is added into the DNA and is repaired with the new DNA that was reverse transcribed (Godbout & Tremblay, 2023). The desired correction needs to be represented within the RTT sequence. Godbout & Tremblay (2023) discuss the accomplishments of prime editing when attempted on, eyes, skin, cancer, cystic fibrosis, liver, and even hereditary diseases. They mention that PE3 (a variation of prime editing), is more efficient than PE2 as it is proved to be more safer and efficient. When PE3 was used for Tay-Sachs and liver disease studies, for example, results seemed to show that there were no off-target mutations. However, when PE3 was used for Beta-

thalassemia and cancer studies there were a few off target mutations (Godbout & Tremblay, 2023). Godbout & Tremblay (2023) address that off-target mutations patterns should be observed. Some prime editing studies have been effective when tested on animals while in humans, prime editing was less efficient (Godbout & Tremblay, 2023). There still seems to be an uncertainty with the results of prime editing. The potential of prime editing treating many conditions is definitely there, but the results varied depending on how the components of prime editing was delivered. Godbout & Tremblay (2023) do agree that prime editing is generally more efficient than CRISPR/Cas 9. Along side the discovery of Prime editing, germline genome editing is discovered. Genome editing can make alterations, additions, deletions that are very precise. Germline editing refers to the edit of a genome which allows for the change to become heritable. The germ cells, gametes, zygotes, and embryos themselves go through genetic alterations (Schweikart, 2021). The role of CRISPR is mentioned as being a sufficient tool which helps create mutations on a chromosome. This allows for a gene drive to copy the mutation for the partner chromosome. Therefore, allowing for future generations to inherit the edited genome. Schweikart (2021) highly focuses on the ethicality of this process and mentioned the unknown when it comes to the success rates. Godbout & Tremblay (2023) validate and highlight the process of PE3 and its preference over CRISPR, but they do mention some faults and what should be done in the future. Additionally, PE3 was seen as more affective than PE2 when compared to each other (Godbout & Tremblay, 2023). Schweikart (2021) does bring up a comparison germline genome editing and somatic genome editing. There is emphasis on how germline is riskier. Somatic genome editing edits individual’s cells instead of germ cells. They are used for therapeutic treatments, but there are concerns of how they can be “enhancements”. So there is this general concern of the two

however, one is more riskier than the other. There is a question of autonomy, fairness, equity, and human rights when addressing germline genome editing in particular. Although its main goal is to help prevent genetic disorders, the risk of off-target mutations occurring, especially when it comes to future generations, is of concern (Schweikart, 2021). This is also emphasized and acknowledged by Godbout & Tremblay (2023). Gene Doping is said to potentially improve athletic performances with athletes. So there is a use of genes or cells in a non therapeutic way and more so using genetic modifying tech. This would be considered as somatic genome editing which means that gene doping does involve the use of CRISPR but on individual’s cells like mentioned previously (Costa, 2021). CRISPR is mentioned to be faulty and unreliable as cells that are altered may be missing anti-cancer mechanisms. CRISPR can also cause Off-target genetic errors which implies that gene doping itself can go wrong (Costa, 2021). The concerns seem to be similar and prevalent in researches like Godbout & Tremblay (2023) and Schweikart (2021); the focus on a possible off-target mutation is emphasized. Costa (2021) mentions that the World Anti-Doping Agency (WADA) has enforced restrictions on gene doping, and are encouraged to completely ban it. There is an issue on abuse with gene doping as it can be extremely difficult to detect, but WADA has been raising funds in order to efficiently identify the use of it (Costa, 2021). Costa (2021) focuses on the future regulations and addresses the lack of specificity with WADA’s regulations. There seems to be an emphasis on stricter and specific guidelines. Especially since many athletes may claim that they are using therapeutic gene therapy, which can be true, but regardless, the gene therapy may be enhancing their skills (Costa, 2021). When the focus of ethicality and gene doping come into question, the focus is on fairness when it comes to sports. There seems to be a need for stricter regulations; not entirely due to mutations and potential risk factors like Godbout

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& Tremblay and Schweikart focus on. These regulations are encouraged in order to prevent the gene doping athletes to compete and offer a disadvantage to other athletes to work hard, which is what Costa emphasizes (2021). There has been major concerns about Gene-edited babies, very similar to the germline genome editing concern. Wang & Yang (2019) address that even if there has been some way to figure out how to effectively avoid or detect off-target mutations, loop holes may occur. On-target mutagenesis effects are an example. These consists of significantly large chromosome deletions and truncations. Additionally, the off-target mutations may occur at ow frequency making it hard to detect (Wang & Yang, 2019). This was concluded based off an experiment that edited the CCR5 gene in order to prevent HIV infections in a set of twins. There was a use of whole- genome sequencing (WGS) to detect the off-target mutations, however, Wang and Yang (2019), like stated above, addressed why the detections could not be accurate. They address the need for additional clear and strict laws, as well as, additional improvement and research for genetic technologies (Wang & Yang 2019). This goes hand in hand with the research from Costa and Schweikart as they both discuss the regulations that need to take place. However, Schweikart (2021) focuses more on human rights legislations. The consequences of human genetic engineering plays a large roll in the ethics and fight for legislation measures. CRISPR could be an issue as there are no determined consequences of certain gene editing. Even with reference to germline modification, it may be difficult to identify the future consequences down the generational line (Fenech, 2018). There needs to be additional research and especially a decrease in the 60% frequency of off-target mutations. Fenech addresses regulations as well and the issues regarding it (very similar to the previous researchers). Human genetic engineering is currently being privately funded which makes it

more difficult for there to be any federal restrictions as federal funding is not in use (Fenech 2018). However, if the states take action, for example, within the U.S. then this can be addressed and clear and strict restrictions may occur. There is a drastic focus and emphasis on the potential harmful and unknown consequences, as well as, regulation. Fenech (2018) is able to bring attention that states within the U.S. can help officially establish strict regulations. While Godbout & Tremblay, Casta, and Wang & Yang, address what other organizations should take into consideration when thinking about regulations and what issues the regulations in general should address. Researchers addressing the issue of germline modifications, emphasized the unknown consequences and mutations that could occur for the child or the future generations. There was not really any information about how this gene editing could be abused to create (designer babies) and inequity. Schweikart (2021) does on the other hand discuss and emphasize human rights, for example, consent and inequity. Researches focused on Somatic gene modifications did address equity and fairness to an extent especially with gene doping. There’s a larger discussion and impact that needs to be addressed on a wider scale. The ethics discussion has to proceed as there are many other factors that contribute to the concerns of human genetic engineering; they can be social, political, or moral. References Costa, S. (2021). The Current State of Gene Doping and the Future of Gene Therapy Regulations. San Diego International Law Journal, 23(1), 157–193.

Fenech, L. (2018). Creating the Perfect Human Race: How Far Will We Go for Designer Families? Family Court Review, 56(1), 150–164. https://doi-org.ccny- proxy1.libr.ccny.cuny.edu/10.1111/fcre.12328 Godbout, K., & Tremblay, J. P. (2023, February 7). Prime editing for human gene therapy: Where are we now? MDPI. Retrieved March 14, 2023, from https://www.mdpi.com/2073- 4409/12/4/536 Schweikart, S. J. (2021). Global Regulation of Germline Genome Editing: Ethical Considerations and Application of International Human Rights Law. Loyola of Los Angeles International & Comparative Law Review, 43(3), 279–296. Wang, H., & Yang, H. (2019). Gene-edited babies: What went wrong and what could go wrong. PLoS Biology, 17(4), 1–5. https://doi-org.ccny-proxy1.libr.ccny.cuny.edu/10.1371/journal.pbio.3000224

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