Bioscience Methods 2024, Vol.15, No.4, 162-172 http://bioscipublisher.com/index.php/bm 1 66 powerful tool for eliminating PERVs and enhancing the biosafety of transplantable pig organs, careful consideration of the associated challenges and ongoing research are essential to fully harness its potential. Figure 2 Detection of PERV expression in different cell lines by immunofluorescence (Adopted from Godehardt et al., 2019) Image caption: A-D, Immunohistology of PERV protein using an anti-PERV nucleocapsid p10 directed antibody26. E-H, represent the p10 pre-serum control. A, E, PERV-A, PERV-B positive PK15 cells, (B, F) PERV-A, PERV-B inactivated PK15 clone 15 cells, (C, G) PERV-B positive 293T/B(33) and (D, H) PERV-negative 293T cells. Viral nucleocapsid p10 is indicated by arrow (Adopted from Godehardt et al., 2019) 4 Enhancing Resistance to Viral and Bacterial Infections 4.1 Genetic modifications to confer resistance to common viral infections (e.g., PRRSV, PCV2) Genetic modifications have shown significant promise in conferring resistance to common viral infections in pigs, particularly Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). One of the most notable strategies involves the modification of the CD163 gene, which encodes a scavenger receptor critical for PRRSV entry into host cells. Several studies have demonstrated that altering or knocking out specific domains of CD163 can confer resistance to PRRSV. For instance, substituting exon 7 of the porcine CD163 gene with the corresponding exon from human CD163-like 1 (hCD163L1) using CRISPR/Cas9 technology has been shown to inhibit PRRSV replication significantly. This modification resulted in a decreased viral load and improved survival rates in infected pigs (Chen et al., 2019). Similarly, complete knockout of the CD163 gene using CRISPR/Cas9 and somatic cell nuclear transfer (SCNT) technologies has rendered pigs fully resistant to highly pathogenic PRRSV, with no adverse effects on macrophage function (Yang et al., 2018). Another approach involved deleting the
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