Molecular genetic status of pigs of ukrainian breeds suitable for use in xenotranplantation
DOI:
https://doi.org/10.31548/dopovidi.2(108).2024.003Keywords:
xenotransplantation, domestic pig, DNA typing, retroviruses, PERV-A, PERV-CAbstract
Thousands of people in Ukraine and the world need organ transplants. However, the main obstacle to the wider use of transplantation remains the shortage of donor organs and tissues. To prevent human infection with viruses during organ transplantation, pigs are carefully screened for PERV retroviruses and animals with low expression levels of PERV-A and PERV-C are selected.
The article is devoted to the discussion of the results of research on the frequency of retrovirus PERV types A and C in populations of Ukrainian pig breeds.
The research was carried out on samples of pigs of the following breeds: Mirgorod, Great White, Poltava meat, Ukrainian meat, Ukrainian Steppe Ryaba, Landras, Pietren, Vietnamese Vyslobryukha, Vietnamese Meishan, and wild pig. Biomaterial samples (venous blood, bristles with hair bulbs) were collected for molecular genetic analysis. Isolation of genomic DNA from the samples was carried out by the salt method and using the Chelex-100 ion exchange resin. Genotyping was performed by the method of allele-specific (PCR-SSP) multiplex polymerase chain reaction using primers complementary to regions of the PERV-C, PERV-A gene loci. A fragment of the domestic pig alpha-actin locus (α-Actin) was used as an internal PCR control. Amplification was carried out in a thermal cycler "Tertsik-2" ("DNA-technology", RF). Electrophoretic separation of amplified DNA sections in multiplex PCR format was performed in a 2% agarose gel in a tris-borate electrophoresis buffer.
It was established that among the studied groups of pigs, the highest concentration of retrovirus PEVR-C was found in animals of the breeds Vietnamese Vyslobrukha (100%), Poltava meat (75%), Ukrainian steppe spotted (55%), large white (50%), landrace (50%). The largest share of PEVR-A retrovirus carrier animals is characterized by the breeds of Vietnamese Vyslobrukha (100%), Poltava meat 95%), Pietren (80%) and Ukrainian meat (73%). The largest number of individuals free from PERV-A and PERV-C retroviruses were found in experimental groups of breeds of Ukrainian steppe spotted (75 and 45%, respectively), Myrhorod (32 and 75%, respectively), landrace (75 and 50%, respectively) and wild pig. Pigs these breeds can become promising organ donors for xenotransplantation.
References
Zhang Y., Xing X., Huang L. et al. (2020). Screening pigs for xenotransplantation in China: investigation of porcine endogenous retrovirus in Diannan small-eared pigs. Virus Genes. 56: 202–208. https://doi.org/10.1007/s11262-019-01722-7
Voloshchuk V.M. et al. (2014). Swine breeding. Monograph. Kyiv, Agrarian science. 587 p
Kaulitz D. Mihica D. Adlhoch C. Semaan M. Denner J. (2013). Improved pig donor screening including newly identified variants of porcine endogenous retrovirus-C (PERV-C). Arch. Virol. 158:341–348 .
Dieckhoff B., Kessler B., Jobst D., Kues W., Petersen B., Pfeifer A., Kurth R., Niemann H., Wolf E., Denner J. (2009). Distribution and expression of porcine endogenous retroviruses in multi-transgenic pigs generated for xenotransplantation. Xenotransplantation. 16: 64–73.
Peacall R, Smouse P.E. (2006). GENALEX 6: genetican alisys in Excel Population genetics of twarean dresearch. Molecul Ecology Notes. 6: 288-295.
Nikitin S.V., Yudin N.S, Knyazev S.P. (2010). Differentiation of wild boar and domestic pig populations based on the frequency of chromosomes carrying endogenous retroviruses. Natural Science. 2(6):527-534. DOI:10.4236/ns.2010.26066.
Godehardt A.W., Fischer N., Rauch P., Gulich B., Boller K., Church G.M., Tönjes R. (2020). Characterization of porcine endogenous retrovirus particles released by the CRISPR/Cas9 inactivated cell line PK15 clone 15. Xenotransplantation. 27( 2): 12563. doi: 10.1111/xen.12563
Denner J. (2021). Porcine Endogenous Retroviruses and Xenotransplantation, Viruses. 13(11): 2156. https://doi.org/10.3390/v13112156
Liu Y., Niu Y., Ma X. et al. (2023). Porcine endogenous retrovirus: classification, molecular structure, regulation, function, and potential risk in xenotransplantation. Funct Integr Genomics 23:60. https://doi.org/10.1007/s10142-023-00984-7
Downloads
Published
Issue
Section
License
Relationship between right holders and users shall be governed by the terms of the license Creative Commons Attribution – non-commercial – Distribution On Same Conditions 4.0 international (CC BY-NC-SA 4.0):https://creativecommons.org/licenses/by-nc-sa/4.0/deed.uk
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
