{"id":227593,"date":"2025-12-21T13:22:26","date_gmt":"2025-12-21T19:22:26","guid":{"rendered":"https:\/\/lifeboat.com\/blog\/2025\/12\/efficient-site-specific-integration-of-kilobase-length-dna-fragments-in-plant-cells-via-kp03-recombinase"},"modified":"2025-12-21T13:22:26","modified_gmt":"2025-12-21T19:22:26","slug":"efficient-site-specific-integration-of-kilobase-length-dna-fragments-in-plant-cells-via-kp03-recombinase","status":"publish","type":"post","link":"https:\/\/lifeboat.com\/blog\/2025\/12\/efficient-site-specific-integration-of-kilobase-length-dna-fragments-in-plant-cells-via-kp03-recombinase","title":{"rendered":"Efficient site-specific integration of kilobase-length DNA fragments in plant cells via Kp03 recombinase"},"content":{"rendered":"

<\/a><\/p>\n

Employing this sensitive assay, we revealed that the integration efficiencies of the 9.9-kb attP-containing DNA donor in rice cells remained remarkably high at 80.5%, relative to the 3.4-kb attP-containing donor, which was set as the baseline for 100% integration efficiency (Figure 2 <\/a>B). However, a significant decrease in recombination efficiency was observed for donor DNA exceeding 17 kb. The integration efficiency of the 17.4-kb attP-containing plasmid decreased to 42.5% (Figure 2 <\/a>B), and for the 27.3-kb attP-containing plasmid, it further declined to 8.2% (Figure 2 <\/a>B). Similar trends were observed in Arabidopsis<\/i>, with recombination efficiencies of 51.4%, 26.5%, and 9.0% for the 9.9-, 17.4-, and 27.3-kb attP-containing donors, respectively (Figure 2 <\/a>C). Furthermore, PCR amplification of the attL junctions confirmed that the donor attP sequences had indeed been recombined into the attB as expected (Figures 2 <\/a>D and 2E). Collectively, these results demonstrate that the Kp03 system can efficiently mediate targeted insertion of large DNA donors up to 27.3 kb in plant cells.<\/p>\n

Interestingly, in addition to donor size, we also observed that Kp03-mediated recombination efficiency is sensitive to temperature, exhibiting differential efficiency under varying thermal conditions (Figure S2<\/a>).<\/p>\n","protected":false},"excerpt":{"rendered":"

Employing this sensitive assay, we revealed that the integration efficiencies of the 9.9-kb attP-containing DNA donor in rice cells remained remarkably high at 80.5%, relative to the 3.4-kb attP-containing donor, which was set as the baseline for 100% integration efficiency (Figure 2 B). However, a significant decrease in recombination efficiency was observed for donor DNA [\u2026]<\/p>\n","protected":false},"author":662,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[11,1506],"tags":[],"class_list":["post-227593","post","type-post","status-publish","format-standard","hentry","category-biotech-medical","category-food"],"_links":{"self":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/227593","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/users\/662"}],"replies":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/comments?post=227593"}],"version-history":[{"count":0,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/227593\/revisions"}],"wp:attachment":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/media?parent=227593"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/categories?post=227593"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/tags?post=227593"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}