Sperm-mediated gene transfer (SMGT) might become the most efficient and cost effective technique to generate transgenic\u00a0animals, which will significantly increase their application in biomedical research and in commercial production. Despite some\u00a0successes, the technique has remained controversial for almost 20 years and despite number of studies the reasons for poor\u00a0reproducibility of this promising technology has not been understood. We suggest that the reason for poor reproducibility is the\u00a0presence of natural defences against exogenous DNA invasion acting in spermatozoa or in embryo. Based on previous reports we\u00a0have investigated the effect of foreign DNA binding on spermatozoa by monitoring motility, viability and genomic DNA damage.\u00a0Evaluation of DNA binding in sperm collected from 16 boars demonstrated that 28\u201345% of the added pEGFP plasmid was bound to\u00a0spermatozoa with 9\u201332% being internalized in sperm nucleus. In agreement with previous reports, our results demonstrated that the\u00a0pEGFP-treated sperm show an average a 2-fold decrease in motility ( p < 0.05), 5-fold decrease in progressive motility ( p < 0.05),\u00a0and 1.4-fold increase in number of sperm with highly damaged DNA ( p < 0.05) as detected by Comet assay. In contrast with\u00a0previous reports, we demonstrate that all such changes were associated with the removal of seminal plasma during the washing step\u00a0and not with foreign DNA binding per se. We suggest that poor reproducibility of SMGT most likely result from selection against\u00a0DNA-loaded sperm at later stages of fertilization.<\/p>\n","protected":false},"excerpt":{"rendered":"
Sperm-mediated gene transfer (SMGT) might become the most efficient and cost effective technique to generate transgenic\u00a0animals, which will significantly increase their application in biomedical research and in commercial production. Despite some\u00a0successes, the technique has remained controversial for almost 20 years and despite number of studies the reasons for poor\u00a0reproducibility of this promising technology has not […]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[18120,19],"tags":[640,5162,190,3734],"class_list":["post-15105","post","type-post","status-publish","format-standard","hentry","category-ontario-pork","category-pork-insight-articles","tag-animal","tag-boar","tag-cost","tag-dna"],"_links":{"self":[{"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/posts\/15105","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/comments?post=15105"}],"version-history":[{"count":1,"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/posts\/15105\/revisions"}],"predecessor-version":[{"id":15107,"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/posts\/15105\/revisions\/15107"}],"wp:attachment":[{"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/media?parent=15105"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/categories?post=15105"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/prairieswine.com\/rsc\/wp-json\/wp\/v2\/tags?post=15105"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}