[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"doc-detail-227614-en":3,"doc-seo-227614-105":30,"detail-sidebar-cat-0-en-105":91},{"code":4,"msg":5,"data":6},0,"success",{"doc_id":7,"user_id":8,"nickname":9,"user_avatar":10,"doc_module":4,"category_id":11,"category_name":12,"doc_title":13,"doc_description":14,"doc_content":15,"file_id":16,"file_url":17,"file_type":18,"file_size":19,"view_count":20,"is_deleted":4,"is_public":20,"is_downloadable":20,"audit_status":20,"page_count":21,"language":22,"language_code":23,"site_id":24,"html_lang":23,"table_of_contents":25,"faqs":26,"seo_title":27,"seo_description":14,"update_tm":28,"read_time":29},227614,8796095027276,"wps_ap_test_251126_0180","https://avatar.qwps.com/avatar/d3BzX2FwX3Rlc3RfMjUxMTI2XzAxODA=",8,"Research & Report","Single-copy insertion of transgenes in Caenorhabditis elegans - Technical report","Single-copy transgene integration in Caenorhabditis elegans is addressed by a new MosSCI strategy that places one transgene into a defined chromosomal site. The approach uses Mos1-mediated mobilization to generate a double-strand break in noncoding DNA, repaired via copying from an extrachromosomal template. Homozygous single-copy insertions can be obtained in under two weeks after injecting roughly 20 worms, supporting endogenous-level expression in both male and female germlines.","TECHNICAL REPORTS  \nSingle-copy insertion of transgenes in Caenorhabditis elegans  \nChristian Frøkjær-Jensen 1,2, M Wayne Davis 1, Christopher E Hopkins 1, Blake J Newman 1, Jason M Thummel 1, Søren-Peter Olesen2, Morten Grunnet2,3 & Erik M Jorgensen 1  \nAt present, transgenes in Caenorhabditis elegans are generated by injecting DNA into the germline. The DNA assembles into asemistable extrachromosomal array composed of many copies of injected DNA. These transgenes are typically overexpressed in somatic cells and silenced in the germline. We have developed a method that inserts a single copy of a transgene into a defined site. Mobilization of a Mos1 transposon generates a double-strand break in noncoding DNA. The break is repaired by copying DNA from an extrachromosomal template into the chromosomal site. Homozygous single-copy insertions can be obtained in less than 2 weeks by injecting approximately 20 worms. We have successfully inserted transgenes as long as 9 kb and verified that single copies are inserted at the targeted site. Single-copy transgenes are expressed at endogenous levels and can be expressed in the female and male germlines.  \nThe method for introducing DNA into C. elegans1 has not changed in the last 17 years and is elegant in its simplicity. DNA injected into the gonad ofa hermaphrodite concatenates to form an extrachromosomal array and is eventually incorporated into the nucleus. Because chromosomes in C. elegans are holocentric in mitosis, any piece of DNA can serve as a centromere, so these extrachromosomal arrays are duplicated and segregated to daughter cells in mitosis. However, this method for generating transgenic lines suffers from several limitations. First, these minichromosomes do not behave like bona fide chromosomes; they are not perfectly stable in mitosis or meiosis. Thus, transgenic animals are mosaic: some cells carry the transgene, whereas others have lost the array. Second, such arrays contain hundreds of copies of the injected DNA, and the genes are overexpressed. This high copy number can cause dominant-negative or toxic effects2. Third, these repetitive arrays are silenced in some tissues, including muscles3,4 and the germline5. The arrays can be silenced even after they are integrated into a chromosome by irradiation, presumably because of transcriptional silencing of arrays6. Finally, arrays change and show ‘drift’ of expression over many generations7,8; drift may arise from changes in the structure of the arrays or by  \nheritable silencing. These limitations complicate studies relying on stable, tissue-specific expression of transgenes.  \nStable changes can be generated at chromosomal sites in rare instances by homologous recombination after biolistic transformation9 or, more effectively, by template-directed repair after excision of a transposon. For example, mobilizing a Tc1 transposon induces a double-strand break at a defined location in a chromosome; the break can be repaired by copying DNA from a transgenic template10,11. The disadvantage of these mutator strains is that there are hundreds of copies of the transposon in the genome; breaks will be induced at many sites, and the frequency of events at any particular site can be quite low. To generate single-copy transposon insertions, the Drosophila Mos1 element was introduced into C. elegans12. It was recently shown that specific DNA changes can be targeted to loci with Mos1 insertions13. This technique, called Mos1 excision-induced transgene-instructed gene conversion (MosTIC), has been used to insert tags or engineer deletions in particular genes. MosTIC relies on the presence of a Mos1 insertion at the genetic locus to be modified. The nematode gene-tagging tools and resources (NemaGENETAG) consortium has generated a large library of Mos1 inserts with known locations in the genome14.  \nIn this study, we adapted intergenic Mos1 elements for the routine insertion of transgenes using a variation of the MosTIC technique called Mos1-m","cbCaiiF7TGOyiVH1","https://ap.wps.com/l/cbCaiiF7TGOyiVH1","pdf",3460588,1,31,"English","en",105,"# Results\n## Insertion method","[{\"question\":\"What problem does the MosSCI method solve compared with traditional DNA injection?\",\"answer\":\"Traditional germline DNA injection forms unstable extrachromosomal arrays, leading to mosaic animals, high copy-number overexpression, and tissue/germline silencing with expression drift. MosSCI enables stable single-copy insertion at a defined chromosomal locus.\"},{\"question\":\"How does MosSCI insert a transgene at a specific chromosomal site?\",\"answer\":\"Mobilization of a Mos1 transposon creates a double-strand break in noncoding DNA, and the break is repaired by copying DNA from an extrachromosomal template into the chromosomal site.\"},{\"question\":\"What expression properties do MosSCI single-copy insertions provide?\",\"answer\":\"Single-copy transgenes are expressed at endogenous levels and can be expressed in both female and male germlines, including tissues that often silence transgenes.\"}]","Single-copy insertion of transgenes in Caenorhabditis elegans - Technical report | PDF",1789010866,78,{"code":4,"msg":31,"data":32},"ok",{"site_id":24,"language":23,"slug":33,"title":13,"keywords":34,"description":14,"schema_data":35,"social_meta":86,"head_meta":88,"extra_data":90,"updated_unix":28},"single-copy-insertion-of-transgenes-in-caenorhabditis-elegans-technical-report","",{"@graph":36,"@context":85},[37,54,68],{"@type":38,"itemListElement":39},"BreadcrumbList",[40,44,48,51],{"item":41,"name":42,"@type":43,"position":20},"https://docshare.wps.com","Home","ListItem",{"item":45,"name":46,"@type":43,"position":47},"https://docshare.wps.com/document/","Document",2,{"item":49,"name":12,"@type":43,"position":50},"https://docshare.wps.com/document/research-report/",3,{"item":52,"name":13,"@type":43,"position":53},"https://docshare.wps.com/document/single-copy-insertion-of-transgenes-in-caenorhabditis-elegans-technical-report/227614/",4,{"url":52,"name":13,"@type":55,"author":56,"headline":13,"publisher":58,"fileFormat":61,"inLanguage":23,"description":14,"dateModified":62,"datePublished":62,"encodingFormat":61,"isAccessibleForFree":63,"interactionStatistic":64},"DigitalDocument",{"name":9,"@type":57},"Person",{"url":41,"name":59,"@type":60},"DocShare","Organization","application/pdf","2026-09-10",true,{"@type":65,"interactionType":66,"userInteractionCount":20},"InteractionCounter",{"@type":67},"ViewAction",{"@type":69,"mainEntity":70},"FAQPage",[71,77,81],{"name":72,"@type":73,"acceptedAnswer":74},"What problem does the MosSCI method solve compared with traditional DNA injection?","Question",{"text":75,"@type":76},"Traditional germline DNA injection forms unstable extrachromosomal arrays, leading to mosaic animals, high copy-number overexpression, and tissue/germline silencing with expression drift. MosSCI enables stable single-copy insertion at a defined chromosomal locus.","Answer",{"name":78,"@type":73,"acceptedAnswer":79},"How does MosSCI insert a transgene at a specific chromosomal site?",{"text":80,"@type":76},"Mobilization of a Mos1 transposon creates a double-strand break in noncoding DNA, and the break is repaired by copying DNA from an extrachromosomal template into the chromosomal site.",{"name":82,"@type":73,"acceptedAnswer":83},"What expression properties do MosSCI single-copy insertions provide?",{"text":84,"@type":76},"Single-copy transgenes are expressed at endogenous levels and can be expressed in both female and male germlines, including tissues that often silence transgenes.","https://schema.org",{"og:url":52,"og:type":87,"og:title":13,"og:site_name":59,"og:description":14},"article",{"robots":89,"canonical":52},"index,follow",{"doc_id":7,"site_id":24},{"code":4,"msg":5,"data":92},[93,97,101,105,110,115,120,123,128,131,135],{"id":20,"doc_module":4,"doc_module_name":46,"category_name":94,"show_sort_weight":95,"slug":96},"Story & Novel",90,"story-novel",{"id":47,"doc_module":4,"doc_module_name":46,"category_name":98,"show_sort_weight":99,"slug":100},"Literature",80,"literature",{"id":53,"doc_module":4,"doc_module_name":46,"category_name":102,"show_sort_weight":103,"slug":104},"Exam",70,"exam",{"id":106,"doc_module":4,"doc_module_name":46,"category_name":107,"show_sort_weight":108,"slug":109},5,"Comic",60,"comic",{"id":111,"doc_module":4,"doc_module_name":46,"category_name":112,"show_sort_weight":113,"slug":114},6,"Technology",50,"technology",{"id":116,"doc_module":4,"doc_module_name":46,"category_name":117,"show_sort_weight":118,"slug":119},7,"Healthcare",40,"healthcare",{"id":11,"doc_module":4,"doc_module_name":46,"category_name":12,"show_sort_weight":121,"slug":122},30,"research-report",{"id":124,"doc_module":4,"doc_module_name":46,"category_name":125,"show_sort_weight":126,"slug":127},9,"Religion & Spirituality",20,"religion-spirituality",{"id":126,"doc_module":4,"doc_module_name":46,"category_name":129,"show_sort_weight":126,"slug":130},"World Cup","world-cup",{"id":132,"doc_module":4,"doc_module_name":46,"category_name":133,"show_sort_weight":132,"slug":134},10,"Lifestyle","lifestyle",{"id":136,"doc_module":4,"doc_module_name":46,"category_name":137,"show_sort_weight":106,"slug":138},19,"General","general"]