{"id":35,"date":"2020-05-28T11:28:06","date_gmt":"2020-05-28T08:28:06","guid":{"rendered":"http:\/\/sites.icgbio.ru\/ontogen\/?page_id=35"},"modified":"2022-08-16T07:26:59","modified_gmt":"2022-08-16T04:26:59","slug":"transgenic-mice","status":"publish","type":"page","link":"https:\/\/sites.icgbio.ru\/ontogen\/transgenic-mice\/","title":{"rendered":"Transgenic mice"},"content":{"rendered":"<p><a href=\"https:\/\/sites.icgbio.ru\/ontogen\/transgenic-mice-ru\/\"><strong><em>\u0422\u0440\u0430\u043d\u0441\u0433\u0435\u043d\u043d\u044b\u0435 \u043c\u044b\u0448\u0438 <\/em><\/strong><\/a><\/p>\n<p>The Department of Molecular Mechanisms of Ontogenesis actively uses novel methods to generate mice with targeted genome modifications. We successfully collaborate with investigators from Institute Cytology and Genetics (Russia, Novosibirsk) and other Russian institutes.<\/p>\n<p>To generate genetically modified mice, we use two main strategies: direct modification of the zygote genome or the chimeras production. With these two approaches we create transgenic mice with random transgene integrations or mice with targeted modifications (using the CRISPR\/Cas system).<\/p>\n<p>Over the past 5 years, more than 20 different lines with various genome modifications have been successfully obtained in our department. At the moment, we have extensive experience in the field of mouse genome modification.<\/p>\n<p>We invite scientific groups for cooperation in the transgenic mice production. Our department has a full cycle for producing genetically-modified mice: from design of targeted modifications and genetic constructs to the generation of genome-modified mouse lines.<\/p>\n<p><strong>Our competences:<\/strong><\/p>\n<ul>\n<li>Design of experiment including CRISPR and genetic constructs;<\/li>\n<\/ul>\n<ul>\n<li>Generation of transgenic mice with random integration sites by traditional pronuclear microinjection;<\/li>\n<li>Generation of knockout mice;<\/li>\n<li>Generation of mice with targeted point mutations;<\/li>\n<li>Generation of knock-in mice;<\/li>\n<li>Generation of humanized mice;<\/li>\n<li>Generation of mice with chromosomal aberrations.<\/li>\n<\/ul>\n<p><strong>For additional information please contact:<\/strong><\/p>\n<p><u><a href=\"http:\/\/assa.bionet.nsc.ru\/en\/person\/65\/\">Nariman Battulin<\/a><\/u><\/p>\n<p>e-mail:<\/p>\n<p><u><a href=\"mailto:battulin@gmail.com\">battulin@gmail.com<\/a><\/u><\/p>\n<p><u><a href=\"mailto:battulin@bionet.nsc.ru\">battulin@bionet.nsc.ru<\/a><\/u><\/p>\n<p>Work phone:<\/p>\n<p>+7 (383) 363-49-63*1110<\/p>\n<p><strong>\u00a0<\/strong><\/p>\n<p><strong>Mouse lines<\/strong><\/p>\n<p><strong>1)\u00a0 \u00a0\u00a0\u00a0 C57BL\/6J-<em>Panx1<\/em><\/strong><strong><sup>em1Koral<\/sup><\/strong><strong>\/Icg \u2013 knockout of <em>Panx1 <\/em>gene by deleting coding exons 3 and 4\u00a0<\/strong><\/p>\n<p>This line was generated in collaboration with Panchin Yuri from Institute for information transmission problems, Russia, Moscow (<u><a href=\"http:\/\/iitp.ru\/ru\/researchlabs\/241.htm\">http:\/\/iitp.ru\/ru\/researchlabs\/241.htm<\/a><\/u>). The mouse line was donated in SPF-vivarium of Institute of Cytology and Genetics, Russia, Novosibirsk (<u><a href=\"http:\/\/spf.bionet.nsc.ru\/\">http:\/\/spf.bionet.nsc.ru\/<\/a><\/u>).<\/p>\n<p>The knockout mouse line was generated by deletion of coding exons 3 and 4 by microinjection of CRISPR\/Cas9 components into fertilized mouse oocytes of C57BL\/6J inbred strain.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Deletion of coding exons 3 and 4 of <em>Panx1 <\/em>gene;<\/li>\n<li>Deletion size &#8212; 3,473 bp;<\/li>\n<li>Genomic coordinates &#8212; chr9: 15006884-15010356 (mm10).<\/li>\n<\/ul>\n<p>References:<\/p>\n<p><strong>2)\u00a0 \u00a0\u00a0\u00a0 C57BL\/6J-<em>Panx1<\/em><\/strong><strong><sup>em2Koral<\/sup><\/strong><strong>\/Icg \u2013 humanization of <em>Panx1<\/em> gene by codon substitution R216H<\/strong><\/p>\n<p>This line was generated in collaboration with Panchin Yuri from Institute for information transmission problems, Russia, Moscow (<u><a href=\"http:\/\/iitp.ru\/ru\/researchlabs\/241.htm\">http:\/\/iitp.ru\/ru\/researchlabs\/241.htm<\/a><\/u>). After, the mouse line was donated in SPF-vivarium of Institute of Cytology and Genetics, Russia, Novosibirsk (<u><a href=\"http:\/\/spf.bionet.nsc.ru\/\">http:\/\/spf.bionet.nsc.ru\/<\/a><\/u>).<\/p>\n<p>The mouse line was generated by HDR-mediated substitution of arginine to histidine in the\u00a0 position 216 of Panx1 gene. Fertilized mouse oocytes of C57BL\/6J inbred strain were injected with a mix of ssODN, Cas9-mRNA and gRNA. This line mimicked human R217H mutation.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Codon substitution R216H (CGG to CAC) in the <em>Panx1 <\/em>gene.<\/li>\n<\/ul>\n<p>References:<\/p>\n<p><strong>3)\u00a0 \u00a0\u00a0\u00a0 C57BL\/6J-<em>H2-Ab1<\/em><\/strong><strong><sup>em1Koral<\/sup><\/strong><strong>\/Icg \u2013 knockout of <em>H2-Ab1<\/em> gene by whole gene deletion<\/strong><\/p>\n<p>The knockout mouse line was generated by whole gene deletion using microinjection of CRISPR components into mouse zygotes.<\/p>\n<p>Homozygotes for targeted null mutations exhibit depletion of mature CD4+ T cells, deficiency in cell-mediated immune responses, and increased susceptibility to viral infections.<\/p>\n<p>This mouse line was donated in SPF-vivarium of Institute of Cytology and Genetics, Russia, Novosibirsk (<u><a href=\"http:\/\/spf.bionet.nsc.ru\/\">http:\/\/spf.bionet.nsc.ru\/<\/a><\/u>).<\/p>\n<p>Two independent mouse lines, C57BL\/6J-<em>H2-Ab1<\/em><sup>em2Koral<\/sup>\/Icg and C57BL\/6J-<em>H2-Ab1<\/em><sup>em3Koral<\/sup>\/Icg, have similar genetic characteristics.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Full deletion of <em>H2-Ab1<\/em> gene;<\/li>\n<li>Deletion size &#8212; 5.8 kb;<\/li>\n<li>Genome coordinates &#8212; hr17: 34263304-34269093 (mm10).<\/li>\n<\/ul>\n<p>References:<\/p>\n<p><strong>4)\u00a0 \u00a0\u00a0\u00a0 C57BL\/6J-<em>Pcbp1<\/em><\/strong><strong><sup>em1Koral<\/sup><\/strong><strong>\/Icg \u2013 knockout of <em>Pcbp1 <\/em>gene\u00a0<\/strong><\/p>\n<p>This line was generated in collaboration with Alexey Tomilin from Institute of Cytology, Saint-Petersburg, Russia, (<u><a href=\"https:\/\/www.incras.ru\/institut\/nauchnye-podrazdelenija\/laboratorija-molekuljarnoj-biologii-stvolovyh-kletok\/\">https:\/\/www.incras.ru\/institut\/nauchnye-podrazdelenija\/laboratorija-molekuljarnoj-biologii-stvolovyh-kletok\/<\/a><\/u>). This line is stored in cryoconservation status in our department.<\/p>\n<p>Mice heterozygous for a knock-out allele exhibit reduced body weight. Mice homozygous for the allele exhibit embryonic lethality between E3.5 and E8.5.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>47 bp deletion including the ATG codon in the first exon of <em>Pcbp1 <\/em>gene<\/li>\n<li>Genomic coordinates &#8212; chr6: 86525884-86525930 (mm10)<\/li>\n<\/ul>\n<p>References:<\/p>\n<p><strong>\u00a05)\u00a0 \u00a0\u00a0\u00a0 C57BL\/6J-<em>Pcbp1<\/em><\/strong><strong><sup>em2Koral<\/sup><\/strong><strong>\/Icg \u2013\u00a0 knockout of <em>Pcbp1 <\/em>gene<\/strong><\/p>\n<p>This line was generated in collaboration with Alexey Tomilin from Institute of Cytology,\u00a0 Saint-Petersburg, Russia,(<u><a href=\"https:\/\/www.incras.ru\/institut\/nauchnye-podrazdelenija\/laboratorija-molekuljarnoj-biologii-stvolovyh-kletok\/\">https:\/\/www.incras.ru\/institut\/nauchnye-podrazdelenija\/laboratorija-molekuljarnoj-biologii-stvolovyh-kletok\/<\/a><\/u>). This line is stored in cryoconservation status in our department.<\/p>\n<p>Mice heterozygous for a knock-out allele exhibit reduced body weight. Mice homozygous for the allele exhibit embryonic lethality between E3.5 and E8.5.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>38 bp insertion in first exon after the 4th codon of Pcbp1 gene;<\/li>\n<li>Genomic coordinates &#8212; chr6: 86525990 (mm10).<\/li>\n<\/ul>\n<p>References:<\/p>\n<p><strong>6)\u00a0 \u00a0\u00a0\u00a0 C57BL\/6J-<em>Cntn6<\/em><\/strong><strong><sup>em1Koral<\/sup><\/strong><strong>\/Icg \u2013 1.137 Mb deletion of <em>Cntn6 <\/em>gene\u00a0<\/strong><\/p>\n<p>This line was generated by full-length gene deletion including 3\u2019 and 5\u2019 regulatory regions. To generate targeted deletion, zygotes from C57BL\/6J inbred strain were injected with a mix of Cas9-mRNA, two gRNA and ssODN.<\/p>\n<p>This mouse line was donated in SPF-vivarium of Institute of Cytology and Genetics, Russia, Novosibirsk (<u><a href=\"http:\/\/spf.bionet.nsc.ru\/\">http:\/\/spf.bionet.nsc.ru\/<\/a><\/u>).<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Full gene deletion including 3\u2019 and 5\u2019 regulatory regions;<\/li>\n<li>Deletion size &#8212; 1.137 Mb;<\/li>\n<li>Genomic coordinates &#8212; chr6: 103842569-104979807 (mm10).<\/li>\n<\/ul>\n<p>Six independent mouse lines have similar genetic characteristics and are stored in the cryo-archive of our department.<\/p>\n<p>References:<\/p>\n<ol>\n<li>Korablev, A.N., Serova, I.A. &amp; Serov, O.L. Generation of megabase-scale deletions, inversions and duplications involving the Contactin-6 gene in mice by CRISPR\/Cas9 technology. BMC Genet 18, 112 (2017).<u><a href=\"https:\/\/doi.org\/10.1186\/s12863-017-0582-7\"> https:\/\/doi.org\/10.1186\/s12863-017-0582-7<\/a><\/u><\/li>\n<li>Pristyazhnyuk, I.E., Minina, J., Korablev, A. et al. Time origin and structural analysis of the induced CRISPR\/cas9 megabase-sized deletions and duplications involving the Cntn6 gene in mice. Sci Rep 9, 14161 (2019). https:\/\/doi.org\/10.1038\/s41598-019-50649-4<\/li>\n<\/ol>\n<p><strong>7)\u00a0 \u00a0\u00a0\u00a0 B6.Cg-<em>Cntn6<\/em><\/strong><strong><sup>em2Koral<\/sup><\/strong><strong>\/Icg \u2013 1.137 Mb duplication of <em>Cntn6 <\/em>gene\u00a0<\/strong><\/p>\n<p>This line was generated by duplication of Cntn6 gene including 3\u2019 and 5\u2019 regulatory regions. To generate duplication, zygotes from C57BL\/6J x CBA hybrid strain were injected with a mix of CRISPR components. Then, each mouse line was backcrossed on C57BL\/6J inbred strain for more than 5 generations. As a result, we obtained two independent mouse lines.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Duplication of <em>Cntn6 <\/em>gene including 3\u2019 and 5\u2019 regulatory regions;<\/li>\n<li>Size of duplication &#8212; 1.137 Mb;<\/li>\n<li>Genomic coordinates chr6: 103842569-104979807 (mm10).<\/li>\n<\/ul>\n<p>One independent mouse line has similar genetic characteristics and is stored in the cryo archive of our department.<\/p>\n<p>References:<\/p>\n<ol>\n<li>Korablev, A.N., Serova, I.A. &amp; Serov, O.L. Generation of megabase-scale deletions, inversions and duplications involving the Contactin-6 gene in mice by CRISPR\/Cas9 technology. BMC Genet 18, 112 (2017).<u><a href=\"https:\/\/doi.org\/10.1186\/s12863-017-0582-7\"> https:\/\/doi.org\/10.1186\/s12863-017-0582-7<\/a><\/u><\/li>\n<li>Pristyazhnyuk, I.E., Minina, J., Korablev, A. et al. Time origin and structural analysis of the induced CRISPR\/cas9 megabase-sized deletions and duplications involving the Cntn6 gene in mice. Sci Rep 9, 14161 (2019). <u><a href=\"https:\/\/doi.org\/10.1038\/s41598-019-50649-4\">https:\/\/doi.org\/10.1038\/s41598-019-50649-4<\/a><\/u><\/li>\n<\/ol>\n<p><strong>8)\u00a0 B6.Cg-<em>Cntn6<\/em><\/strong><strong><sup>em3Koral<\/sup><\/strong><strong>\/Icg \u2013 1.137 Mb inversion of <em>Cntn6<\/em> gene\u00a0<\/strong><\/p>\n<p>This line was generated by full-length gene inversion including 3\u2019 and 5\u2019 regulatory regions. To generate inversion, zygotes from C57BL\/6J x CBA hybrid strain were injected with a mix of CRISPR components. Then, each mouse line was backcrossed on C57BL\/6J inbred strain for more than 5 generations. As a result, we obtained two independent mouse lines.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Inversion of <em>Cntn6 <\/em>including 3\u2019 and 5\u2019 regulatory regions;<\/li>\n<li>Size of inversion &#8212; 1.137 Mb;<\/li>\n<li>Genomic coordinates &#8212; chr6: 103842569-104979807 (mm10).<\/li>\n<\/ul>\n<p>One independent mouse line has similar genetic characteristics and is stored in the cryo-archive of our department.<\/p>\n<p>References:<\/p>\n<ol>\n<li>Korablev, A.N., Serova, I.A. &amp; Serov, O.L. Generation of megabase-scale deletions, inversions and duplications involving the Contactin-6 gene in mice by CRISPR\/Cas9 technology. BMC Genet 18, 112 (2017).<u><a href=\"https:\/\/doi.org\/10.1186\/s12863-017-0582-7\"> https:\/\/doi.org\/10.1186\/s12863-017-0582-7<\/a><\/u><\/li>\n<\/ol>\n<p><strong>9)\u00a0 C57BL\/6J-<em>Tnf<\/em><\/strong><strong><sup>em1Koral<\/sup><\/strong><strong>\/Icg \u2013 knockout of <em>Tnf-\u03b1 <\/em>gene<\/strong><\/p>\n<p>The knockout mouse line was generated by a 16 bp insertion, introducing a frameshift mutation in the first exon. To generate knockout mouse strain, zygotes from C57BL\/6J inbred strain were injected with a mix of Cas9-protein, gRNA and ssODN. This line was generated in collaboration with\u00a0 <a href=\"http:\/\/assa.bionet.nsc.ru\/en\/person\/53\/\">Darya Bazovkina<\/a> from Institute of Cytology and Genetics, Russia, Novosibirsk.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>16 bp insertion in the first exon, leading to nonsense frameshift mutation.<\/li>\n<\/ul>\n<p>References:<\/p>\n<p><strong>12)\u00a0 C57BL\/6J-<em>Ptpn5<\/em><\/strong><strong><sup>em1Koral<\/sup><\/strong><strong>\/Icg \u2013 knockout of <em>Ptpn5 <\/em>gene<\/strong><\/p>\n<p>The knockout mouse line was generated by microinjection of CRISPR components into C57BL\/6J mouse zygotes leading to deletion of the 12-13 exons. This line was generated in collaboration with <u><a href=\"http:\/\/assa.bionet.nsc.ru\/en\/person\/948\/\">Elizaveta Kulikova<\/a><\/u> from Institute of Cytology and Genetics, Russia, Novosibirsk.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Deletion of exon 12 (partial) and exon 13 (full), which affected Ptpn5 substrate-binding domain;<\/li>\n<li>Deletion size &#8212; 347 bp;<\/li>\n<li>Genome coordinates &#8212; chr7: 47079194-47079540 (mm10).<\/li>\n<\/ul>\n<p>References:<\/p>\n<p><strong>10)\u00a0 B6.Cg-<em>Kit<\/em><\/strong><strong><sup>em1Koral<\/sup><\/strong><strong>\/Icg &#8212; knockout of the <em>Kit <\/em>gene including deletion of a TAD border<\/strong><\/p>\n<p>This line was generated by microinjection of CRISPR components into mouse zygotes. Spontaneous deletion was formed at the site of CRISPR-induced DSB.<\/p>\n<p>To generate modification, zygotes from C57BL\/6J x CBA hybrid strain were injected with a mix of CRISPR components. Each mouse line was backcrossed on C57BL\/6J inbred strain for more than 5 generations.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Deletion of exons 2-21 of the <em>Kit <\/em>gene including the TAD border between the <em>Kit<\/em> locus and the <em>Kdr <\/em>locus. Only the first exon coding for the KIT signal peptide remained intact (aa sequence MRGARGAWDLLCVLLVLLRGQT);<\/li>\n<li>Deletion size &#8212; 293 kb;<\/li>\n<li>Genomic coordinates &#8212; chr5: 75588218-75881214 (mm10).<\/li>\n<\/ul>\n<p>Phenotype:<\/p>\n<ul>\n<li>Mutations at this locus affect migration of embryonic stem cell populations, resulting in mild to severe impairments in hematopoiesis and pigmentation. While wild-type mice have black coats, heterozygous mutants demonstrate additional abdominal white spots.<\/li>\n<\/ul>\n<p>References:<\/p>\n<ol start=\"2\">\n<li>Korablev, A.; Lukyanchikova, V.; Serova, I.; Battulin, N. On-Target CRISPR\/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes. <em> J. Mol. Sci.<\/em> <strong>2020<\/strong>, <em>21<\/em>, 3604. <u><a href=\"https:\/\/doi.org\/10.3390\/ijms21103604\">https:\/\/doi.org\/10.3390\/ijms21103604<\/a><\/u><\/li>\n<\/ol>\n<p><strong>11) B6;129P-<em>Csn1s1<\/em><\/strong><strong><sup>tm1Icg<\/sup><\/strong><strong>\/Icg \u2013 insertion of the human GMCSF gene after the promoter of the endogenous mouse Csn1s1 (\u03b1-S1-casein) gene<\/strong><\/p>\n<p>The mouse line was created by injections of transgenic 129S2\/SvPasCrl mESCs into blastocysts resulting in chimeric animals. Experiments were performed by Dr. Galina Kontsevaya at SPF-vivarium of Institute of Cytology and Genetics, Novosibirsk, Russia.<\/p>\n<p>Genetics:<\/p>\n<ul>\n<li>Insertion of the human <em>GMCSF<\/em> gene (2029 bp) into the mouse <em>Csn1s1<\/em> gene ORF, right after the start codon.<\/li>\n<li>Genomic coordinates &#8212; chr5: 87667246 (mm10).<\/li>\n<\/ul>\n<p>Phenotype:<\/p>\n<ul>\n<li>Pups from homozygous females grow slower due to the lack of \u03b1-S1-casein in the milk of lactating females.<\/li>\n<li>Milk from transgenic females contains 2-5 ug\/ml hGMCSF.<\/li>\n<\/ul>\n<p><strong>\u00a012) B6;129P-Csn1s1<\/strong><strong><sup>tm2Icg<\/sup><\/strong><strong>\/Icg \u2013 replacement of the mouse <em>Csn1s1<\/em> (\u03b1-S1-casein) gene with the human<em> GMCSF<\/em> gene<\/strong><\/p>\n<p>The mouse line was created by injections of transgenic 129S2\/SvPasCrl mESCs into blastocysts resulting in chimeric animals. Experiments were performed by Dr. Galina Kontsevaya at SPF-vivarium of Institute of Cytology and Genetics, Novosibirsk, Russia.<\/p>\n<p>enetics:<\/p>\n<ul>\n<li>Insertion of the human <em>GMCSF<\/em> gene (2029 bp) into the mouse <em>Csn1s1<\/em> gene ORF, right after the start codon. Concomitantly, the <em>Csn1s1<\/em> coding region (13,7 kbp) was deleted.<\/li>\n<li>Genomic coordinates &#8212; chr5: 87667246-87680963 (mm10).<\/li>\n<\/ul>\n<p>Phenotype:<\/p>\n<ul>\n<li>Pups from homozygous females grow slower due to the lack of \u03b1-S1-casein in the milk of lactating females.<\/li>\n<li>Milk from transgenic females contains 100-500 ug\/ml hGMCSF.<\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"<p>\u0422\u0440\u0430\u043d\u0441\u0433\u0435\u043d\u043d\u044b\u0435 \u043c\u044b\u0448\u0438 The Department of Molecular Mechanisms of Ontogenesis actively uses novel methods to generate mice with targeted genome modifications. We successfully collaborate with investigators from Institute Cytology and Genetics (Russia, Novosibirsk) and other Russian institutes. To generate genetically modified mice, we use two main strategies: direct modification of the zygote genome or the chimeras&#8230;<\/p>\n","protected":false},"author":3,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/sites.icgbio.ru\/ontogen\/wp-json\/wp\/v2\/pages\/35"}],"collection":[{"href":"https:\/\/sites.icgbio.ru\/ontogen\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/sites.icgbio.ru\/ontogen\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/sites.icgbio.ru\/ontogen\/wp-json\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.icgbio.ru\/ontogen\/wp-json\/wp\/v2\/comments?post=35"}],"version-history":[{"count":5,"href":"https:\/\/sites.icgbio.ru\/ontogen\/wp-json\/wp\/v2\/pages\/35\/revisions"}],"predecessor-version":[{"id":68,"href":"https:\/\/sites.icgbio.ru\/ontogen\/wp-json\/wp\/v2\/pages\/35\/revisions\/68"}],"wp:attachment":[{"href":"https:\/\/sites.icgbio.ru\/ontogen\/wp-json\/wp\/v2\/media?parent=35"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}