Bioscience Methods 2024, Vol.15, No.4, 162-172 http://bioscipublisher.com/index.php/bm 1 71 recipients (Ryczek et al., 2020). Furthermore, the identification of PCMV as a critical factor in transplant survival underscores the importance of rigorous screening and elimination of zoonotic pathogens from donor pigs. Ensuring the absence of such pathogens will significantly improve the biosafety of transplantable pig organs and increase the likelihood of successful clinical applications (Denner, 2018). These findings collectively suggest that with continued advancements in genetic engineering and pathogen control, xenotransplantation could become a viable solution to the organ shortage crisis. To fully realize the potential of xenotransplantation, it is imperative to continue research in genetic modifications and pathogen elimination. Researchers should focus on refining CRISPR/Cas9 techniques to further minimize off-target effects and enhance the precision of genetic edits. Additionally, there is a need for comprehensive studies on the transmission mechanisms and impacts of zoonotic pathogens like PCMV to develop effective strategies for their elimination (Denner, 2018). Interdisciplinary collaboration among geneticists, immunologists, virologists, and transplant surgeons is essential to address the multifaceted challenges of xenotransplantation. By working together, these experts can develop innovative solutions that ensure the safety and efficacy of transplantable pig organs, ultimately improving patient outcomes and addressing the global organ shortage. Acknowledgments Authors extends our sincere thanks to two anonymous peer reviewers for their invaluable feedback on the initial draft of this manuscript. Conflict of Interest Disclosure Authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. Reference Burkard C., Opriessnig T., Mileham A., Stadejek T., Ait-Ali T., Lillico S., Whitelaw C., and Archibald A., 2018, Pigs lacking the scavenger receptor cysteine-rich domain 5 of CD163 are resistant to porcine reproductive and respiratory syndrome virus 1 infection, Journal of Virology, 92(16): 10.1128/jvi. 00415-18. https://doi.org/10.1128/JVI.00415-18 Chen J., Wang H., Bai J., Liu W., Liu X., Yu D., Feng T., Sun Z., Zhang L., Ma L., Hu Y., Zou Y., Tan T., Zhong J., Hu M., Bai X., Pan D., Xing Y., Zhao Y., Tian K., Hu X., and Li, N., 2019, Generation of pigs resistant to highly pathogenic-porcine reproductive and respiratory syndrome virus through gene editing of CD163, International Journal of Biological Sciences, 15(2): 481-492 https://doi.org/10.7150/ijbs.25862 Cooper D., Hara H., Iwase H., Yamamoto T., Li Q., Ezzelarab M., Federzoni E., Dandro A., and Ayares D., 2019, Justification of specific genetic modifications in pigs for clinical organ xenotransplantation, Xenotransplantation, 26(4): e12516 https://doi.org/10.1111/xen.12516 Deng J., Yang L., Wang Z., Ouyang H., Yu H., Yuan H., and Pang D., 2022, Advance of genetically modified pigs in xeno-transplantation, Frontiers in Cell and Developmental Biology, 10: 1033197. https://doi.org/10.3389/fcell.2022.1033197 Denner J., 2018, Reduction of the survival time of pig xenotransplants by porcine cytomegalovirus, Virology Journal, 15: 1-7. https://doi.org/10.1186/s12985-018-1088-2 Denner J., 2021, Porcine endogenous retroviruses and xenotransplantation, Viruses, 13(11): 2156. https://doi.org/10.3390/v13112156 Denner J., 2022, Virus safety of xenotransplantation, Viruses, 14(9): 1926. https://doi.org/10.3390/v14091926 Godehardt A., Fischer N., Rauch P., Gulich B., Boller K., Church G., and Tönjes R., 2019, Characterization of porcine endogenous retrovirus particles released by the CRISPR/Cas9 inactivated cell line PK15 clone 15, Xenotransplantation, 27(2): e12563. https://doi.org/10.1111/xen.12563 Hirata M., Wittayarat M., Hirano T., Nguyen N., Le Q., Namula Z., Fahrudin M., Tanihara F., and Otoi T., 2019, The relationship between embryonic development and the efficiency of target mutations in porcine endogenous retroviruses (PERVs) pol genes in porcine embryos, Animals, 9(9): 593. https://doi.org/10.3390/ani9090593 Kemter E., Denner J., and Wolf, E. 2018, Will genetic engineering carry xenotransplantation of pig islets to the clinic, Current Diabetes Reports, 18: 1-12. https://doi.org/10.1007/s11892-018-1074-5 Lei T., Chen L., Wang K., Du S., Gonelle-Gispert C., Wang Y., and Buhler L., 2022, Genetic engineering of pigs for xenotransplantation to overcome immune rejection and physiological incompatibilities: the first clinical steps, Frontiers in Immunology, 13: 1031185. https://doi.org/10.3389/fimmu.2022.1031185
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