Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization
Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by in...
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| Veröffentlicht in: | Scientific reports 2019-08, Vol.9 (1), p.11571-11571, Article 11571 |
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| creator | Honda, Arata Tachibana, Ryoma Hamada, Kazuya Morita, Kohtaro Mizuno, Naoaki Morita, Kento Asano, Masahide |
| description | Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by
in vitro
fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the
tyrosinase
gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3–47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats. |
| doi_str_mv | 10.1038/s41598-019-47964-1 |
| format | Article |
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in vitro
fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the
tyrosinase
gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3–47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-019-47964-1</identifier><identifier>PMID: 31399630</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>13 ; 42/41 ; 45 ; 631/1647/334/1874/486 ; 631/208/135 ; 631/61/17/1511 ; 64 ; Anesthesia ; Animals ; CRISPR-Cas Systems ; Electroporation ; Electroporation - methods ; Electroporation - veterinary ; Embryos ; Female ; Fertilization in Vitro - methods ; Fertilization in Vitro - veterinary ; Gene Editing - methods ; Gene Editing - veterinary ; Gene Knock-In Techniques - methods ; Gene Knock-In Techniques - veterinary ; Gene Knockout Techniques - methods ; Gene Knockout Techniques - veterinary ; Genome editing ; Genomes ; Humanities and Social Sciences ; In vitro fertilization ; Inhibin ; Male ; multidisciplinary ; Offspring ; Oocytes ; Ovulation ; Rats - embryology ; Rats - genetics ; Rats - physiology ; Rats, Inbred F344 - embryology ; Rats, Inbred F344 - genetics ; Rats, Inbred F344 - physiology ; Rats, Long-Evans - embryology ; Rats, Long-Evans - genetics ; Rats, Long-Evans - physiology ; Rats, Sprague-Dawley - embryology ; Rats, Sprague-Dawley - genetics ; Rats, Sprague-Dawley - physiology ; Rats, Wistar - embryology ; Rats, Wistar - genetics ; Rats, Wistar - physiology ; Rodents ; Science ; Science (multidisciplinary) ; Superovulation ; Tyrosinase gene</subject><ispartof>Scientific reports, 2019-08, Vol.9 (1), p.11571-11571, Article 11571</ispartof><rights>The Author(s) 2019</rights><rights>This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c584t-491959e09a3ad491480cfd66bc576bc798f26e4f8a86ab6bc41a8369b74356033</citedby><cites>FETCH-LOGICAL-c584t-491959e09a3ad491480cfd66bc576bc798f26e4f8a86ab6bc41a8369b74356033</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689013/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689013/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27924,27925,41120,42189,51576,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31399630$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Honda, Arata</creatorcontrib><creatorcontrib>Tachibana, Ryoma</creatorcontrib><creatorcontrib>Hamada, Kazuya</creatorcontrib><creatorcontrib>Morita, Kohtaro</creatorcontrib><creatorcontrib>Mizuno, Naoaki</creatorcontrib><creatorcontrib>Morita, Kento</creatorcontrib><creatorcontrib>Asano, Masahide</creatorcontrib><title>Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by
in vitro
fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the
tyrosinase
gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3–47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats.</description><subject>13</subject><subject>42/41</subject><subject>45</subject><subject>631/1647/334/1874/486</subject><subject>631/208/135</subject><subject>631/61/17/1511</subject><subject>64</subject><subject>Anesthesia</subject><subject>Animals</subject><subject>CRISPR-Cas Systems</subject><subject>Electroporation</subject><subject>Electroporation - methods</subject><subject>Electroporation - veterinary</subject><subject>Embryos</subject><subject>Female</subject><subject>Fertilization in Vitro - methods</subject><subject>Fertilization in Vitro - veterinary</subject><subject>Gene Editing - methods</subject><subject>Gene Editing - veterinary</subject><subject>Gene Knock-In Techniques - methods</subject><subject>Gene Knock-In Techniques - veterinary</subject><subject>Gene Knockout Techniques - methods</subject><subject>Gene Knockout Techniques - veterinary</subject><subject>Genome editing</subject><subject>Genomes</subject><subject>Humanities and Social Sciences</subject><subject>In vitro fertilization</subject><subject>Inhibin</subject><subject>Male</subject><subject>multidisciplinary</subject><subject>Offspring</subject><subject>Oocytes</subject><subject>Ovulation</subject><subject>Rats - embryology</subject><subject>Rats - genetics</subject><subject>Rats - physiology</subject><subject>Rats, Inbred F344 - embryology</subject><subject>Rats, Inbred F344 - genetics</subject><subject>Rats, Inbred F344 - physiology</subject><subject>Rats, Long-Evans - embryology</subject><subject>Rats, Long-Evans - genetics</subject><subject>Rats, Long-Evans - physiology</subject><subject>Rats, Sprague-Dawley - embryology</subject><subject>Rats, Sprague-Dawley - genetics</subject><subject>Rats, Sprague-Dawley - physiology</subject><subject>Rats, Wistar - embryology</subject><subject>Rats, Wistar - genetics</subject><subject>Rats, Wistar - physiology</subject><subject>Rodents</subject><subject>Science</subject><subject>Science (multidisciplinary)</subject><subject>Superovulation</subject><subject>Tyrosinase gene</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kctOLCEQhok5Rs3oC7gwJG7OppVb07AxOTHjJTFxo2tCd8OI9oACPcn49DLOeDsLWQBFffUXlR-AQ4xOMKLiNDFcS1EhLCvWSM4qvAX2CGJ1RSghf77dd8FBSo-orJpIhuUO2KWYSskp2gP91FrXOeMz7E10C51d8DBY-ORD91SFMUPt-03kPIw6Jzgm52fQzNu4DAmGNmvnTQ_bJSzEwuUYoDUxu8G9vuvtg22rh2QONucE3F9M786vqpvby-vzfzdVVwuWKyaxrKVBUlPdl4AJ1Nme87arm7I1UljCDbNCC67b8sKwFpTLtmG05ojSCThb6z6P7dz0XZkq6kE9RzfXcamCdupnxrsHNQsLxbmQCK8E_m4EYngZTcpq7lJnhkF7E8akCGmwYM2q2QQc_4c-hjH6Mt6KQjWmrGkKRdZUF0NK0djPz2CkVj6qtY-q-KjefVS4FB19H-Oz5MO1AtA1kErKz0z86v2L7BvWmqms</recordid><startdate>20190809</startdate><enddate>20190809</enddate><creator>Honda, Arata</creator><creator>Tachibana, Ryoma</creator><creator>Hamada, Kazuya</creator><creator>Morita, Kohtaro</creator><creator>Mizuno, Naoaki</creator><creator>Morita, Kento</creator><creator>Asano, Masahide</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>C6C</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20190809</creationdate><title>Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization</title><author>Honda, Arata ; Tachibana, Ryoma ; Hamada, Kazuya ; Morita, Kohtaro ; Mizuno, Naoaki ; Morita, Kento ; Asano, Masahide</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c584t-491959e09a3ad491480cfd66bc576bc798f26e4f8a86ab6bc41a8369b74356033</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>13</topic><topic>42/41</topic><topic>45</topic><topic>631/1647/334/1874/486</topic><topic>631/208/135</topic><topic>631/61/17/1511</topic><topic>64</topic><topic>Anesthesia</topic><topic>Animals</topic><topic>CRISPR-Cas Systems</topic><topic>Electroporation</topic><topic>Electroporation - 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embryology</topic><topic>Rats, Long-Evans - genetics</topic><topic>Rats, Long-Evans - physiology</topic><topic>Rats, Sprague-Dawley - embryology</topic><topic>Rats, Sprague-Dawley - genetics</topic><topic>Rats, Sprague-Dawley - physiology</topic><topic>Rats, Wistar - embryology</topic><topic>Rats, Wistar - genetics</topic><topic>Rats, Wistar - physiology</topic><topic>Rodents</topic><topic>Science</topic><topic>Science (multidisciplinary)</topic><topic>Superovulation</topic><topic>Tyrosinase gene</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Honda, Arata</creatorcontrib><creatorcontrib>Tachibana, Ryoma</creatorcontrib><creatorcontrib>Hamada, Kazuya</creatorcontrib><creatorcontrib>Morita, Kohtaro</creatorcontrib><creatorcontrib>Mizuno, Naoaki</creatorcontrib><creatorcontrib>Morita, Kento</creatorcontrib><creatorcontrib>Asano, Masahide</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Honda, Arata</au><au>Tachibana, Ryoma</au><au>Hamada, Kazuya</au><au>Morita, Kohtaro</au><au>Mizuno, Naoaki</au><au>Morita, Kento</au><au>Asano, Masahide</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2019-08-09</date><risdate>2019</risdate><volume>9</volume><issue>1</issue><spage>11571</spage><epage>11571</epage><pages>11571-11571</pages><artnum>11571</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by
in vitro
fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the
tyrosinase
gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3–47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>31399630</pmid><doi>10.1038/s41598-019-47964-1</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | 13 42/41 45 631/1647/334/1874/486 631/208/135 631/61/17/1511 64 Anesthesia Animals CRISPR-Cas Systems Electroporation Electroporation - methods Electroporation - veterinary Embryos Female Fertilization in Vitro - methods Fertilization in Vitro - veterinary Gene Editing - methods Gene Editing - veterinary Gene Knock-In Techniques - methods Gene Knock-In Techniques - veterinary Gene Knockout Techniques - methods Gene Knockout Techniques - veterinary Genome editing Genomes Humanities and Social Sciences In vitro fertilization Inhibin Male multidisciplinary Offspring Oocytes Ovulation Rats - embryology Rats - genetics Rats - physiology Rats, Inbred F344 - embryology Rats, Inbred F344 - genetics Rats, Inbred F344 - physiology Rats, Long-Evans - embryology Rats, Long-Evans - genetics Rats, Long-Evans - physiology Rats, Sprague-Dawley - embryology Rats, Sprague-Dawley - genetics Rats, Sprague-Dawley - physiology Rats, Wistar - embryology Rats, Wistar - genetics Rats, Wistar - physiology Rodents Science Science (multidisciplinary) Superovulation Tyrosinase gene |
| title | Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization |
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