International Journal of Clinical Case Reports 2024, Vol.14, No.2, 94-106 http://medscipublisher.com/index.php/ijccr 99 Figure 3 Schematic workflow of the generation of triple-knockout Jeju Native Pigs (JNPs), including gene editing strategies and steps required to obtain the modified JNPs (Adopted from Yoon et al., 2022) Image caption: (A) Schematic diagram for triple-knockout development; (B) the sequences of GGTA1, CMAH, and B4GALNT2; (C) deep sequence analysis of the target region of genes in delivered piglets and details of the target sequences in exon 7 (GGTA1), exon 2 (CMAH), and exon 4 (B4GALNT2); (D) knockout piglets produced. SCNT, somatic cell nuclear transfer (Adopted from Yoon et al., 2022). 4 Enhancing Organ Longevity with CRISPR-Cas9 4.1 Genetic modifications aimed at improving organ longevity CRISPR-Cas9 technology has facilitated significant advancements in genetic modifications to enhance organ longevity post-transplantation. Key modifications focus on increasing resistance to ischemia-reperfusion injury (IRI), reducing inflammation, and promoting tissue repair mechanisms. For instance, upregulating the expression of anti-apoptotic genes such as Bcl-2 and heme oxygenase-1 (HO-1) has proven effective in protecting cells from apoptosis and oxidative stress (Wang et al., 2018). Additionally, targeting genes involved in inflammatory pathways, such as NF-κB and its associated cytokines, can mitigate the inflammatory response and subsequent tissue damage (Kurzhagen et al., 2023). Another critical strategy involves modulating antioxidant pathways. Enhancing the expression of genes like Nrf2, which regulates the cellular antioxidant response, has shown potential in reducing oxidative damage and improving organ function (Kurzhagen et al., 2023). 4.2 Mechanisms by which gene editing enhances organ durability Gene editing enhances organ durability through several key mechanisms. One approach involves the upregulation of anti-apoptotic genes, such as Bcl-2, which prevents programmed cell death in transplanted tissues. This strategy is crucial for reducing cell loss during the transplantation and reperfusion phases (Bilbao et al., 1999). Another important mechanism is increasing resistance to ischemia-reperfusion injury. Upregulating genes like HO-1 and using CRISPR to knock out pro-inflammatory genes (e.g., TNF-α) helps to mitigate the effects of IRI by reducing oxidative stress, inflammation, and cellular apoptosis. Modulating genes involved in cellular stress responses, such as Nrf2, also enhances the organ's ability to withstand oxidative stress and inflammation. For instance, CRISPR/Cas9-mediated Keap1 knockout augments Nrf2 activity, boosting the organ's resilience (Kurzhagen et al., 2023). Additionally, gene editing can target
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