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Targeted gene deletion of miRNAs in mice by TALEN system
Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the ap...
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| Published in: | PloS one 2013-10, Vol.8 (10), p.e76004 |
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| creator | Takada, Shuji Sato, Tempei Ito, Yoshiaki Yamashita, Satoshi Kato, Tomoko Kawasumi, Miyuri Kanai-Azuma, Masami Igarashi, Arisa Kato, Tomomi Tamano, Moe Asahara, Hiroshi |
| description | Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the application of TALEN system for mice and successfully obtained gene targeting technique in mice for intergenic region and series of microRNAs. Microinjection of synthesized RNA of TALEN targeting each gene in one cell stage of embryo was carried out and injected oocytes were transferred into pseudopregnant ICR female mice, producing a high success rate of the targeted deletion of miRNA genes. In our condition, TALEN RNA without poly(A) tail worked better than that of with poly(A) tail. This mutated allele in miRNA was transmitted to the next generation, suggesting the successful germ line transmission of this targeting method. Consistent with our notion of miRNAs maturation mechanism, in homozygous mutant mice of miR-10a, the non- mutated strand of miRNAs expression was completely diminished. This method will lead us to expand and accelerate our genetic research using mice in a high throughput way. |
| doi_str_mv | 10.1371/journal.pone.0076004 |
| format | article |
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However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the application of TALEN system for mice and successfully obtained gene targeting technique in mice for intergenic region and series of microRNAs. Microinjection of synthesized RNA of TALEN targeting each gene in one cell stage of embryo was carried out and injected oocytes were transferred into pseudopregnant ICR female mice, producing a high success rate of the targeted deletion of miRNA genes. In our condition, TALEN RNA without poly(A) tail worked better than that of with poly(A) tail. This mutated allele in miRNA was transmitted to the next generation, suggesting the successful germ line transmission of this targeting method. Consistent with our notion of miRNAs maturation mechanism, in homozygous mutant mice of miR-10a, the non- mutated strand of miRNAs expression was completely diminished. This method will lead us to expand and accelerate our genetic research using mice in a high throughput way.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0076004</identifier><identifier>PMID: 24146809</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Animal species ; Animals ; Base Sequence ; Childrens health ; Chromosomes ; Clonal deletion ; Deoxyribonucleases, Type II Site-Specific - genetics ; Deoxyribonucleases, Type II Site-Specific - metabolism ; Deoxyribonucleic acid ; DNA ; DNA, Intergenic - genetics ; DNA, Intergenic - metabolism ; DNA-Binding Proteins - genetics ; DNA-Binding Proteins - metabolism ; Embryo, Mammalian ; Female ; Gene Deletion ; Gene mutation ; Gene sequencing ; Gene targeting ; Gene Targeting - methods ; Genes ; Genetic Engineering ; Genetic modification ; Genetically modified organisms ; Genomes ; House mouse ; Mice ; Mice, Inbred ICR ; Microinjection ; Microinjections ; MicroRNA ; MicroRNAs ; MicroRNAs - genetics ; MicroRNAs - metabolism ; miRNA ; Molecular Sequence Data ; Oocytes ; Oocytes - cytology ; Oocytes - growth & development ; Oocytes - metabolism ; Plasmids ; Poly(A) ; Polyadenine ; Polyadenylation ; Pseudopregnancy ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; Ribonucleic acid ; RNA ; Rodents ; Stem cells ; Transcription activator-like effector nucleases ; Xenopus</subject><ispartof>PloS one, 2013-10, Vol.8 (10), p.e76004</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013 Takada et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013 Takada et al 2013 Takada et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-d019b7181570cca189b320c0f408ca3fe75211c2f5772972e9307030e1ac077a3</citedby><cites>FETCH-LOGICAL-c692t-d019b7181570cca189b320c0f408ca3fe75211c2f5772972e9307030e1ac077a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/1442466781/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/1442466781?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,315,733,786,790,891,25783,27957,27958,37047,44625,53827,53829,75483</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24146809$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Shih, Chiaho</contributor><creatorcontrib>Takada, Shuji</creatorcontrib><creatorcontrib>Sato, Tempei</creatorcontrib><creatorcontrib>Ito, Yoshiaki</creatorcontrib><creatorcontrib>Yamashita, Satoshi</creatorcontrib><creatorcontrib>Kato, Tomoko</creatorcontrib><creatorcontrib>Kawasumi, Miyuri</creatorcontrib><creatorcontrib>Kanai-Azuma, Masami</creatorcontrib><creatorcontrib>Igarashi, Arisa</creatorcontrib><creatorcontrib>Kato, Tomomi</creatorcontrib><creatorcontrib>Tamano, Moe</creatorcontrib><creatorcontrib>Asahara, Hiroshi</creatorcontrib><title>Targeted gene deletion of miRNAs in mice by TALEN system</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the application of TALEN system for mice and successfully obtained gene targeting technique in mice for intergenic region and series of microRNAs. Microinjection of synthesized RNA of TALEN targeting each gene in one cell stage of embryo was carried out and injected oocytes were transferred into pseudopregnant ICR female mice, producing a high success rate of the targeted deletion of miRNA genes. In our condition, TALEN RNA without poly(A) tail worked better than that of with poly(A) tail. This mutated allele in miRNA was transmitted to the next generation, suggesting the successful germ line transmission of this targeting method. Consistent with our notion of miRNAs maturation mechanism, in homozygous mutant mice of miR-10a, the non- mutated strand of miRNAs expression was completely diminished. This method will lead us to expand and accelerate our genetic research using mice in a high throughput way.</description><subject>Animal species</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Childrens health</subject><subject>Chromosomes</subject><subject>Clonal deletion</subject><subject>Deoxyribonucleases, Type II Site-Specific - genetics</subject><subject>Deoxyribonucleases, Type II Site-Specific - metabolism</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Intergenic - genetics</subject><subject>DNA, Intergenic - metabolism</subject><subject>DNA-Binding Proteins - genetics</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Embryo, Mammalian</subject><subject>Female</subject><subject>Gene Deletion</subject><subject>Gene mutation</subject><subject>Gene sequencing</subject><subject>Gene targeting</subject><subject>Gene Targeting - methods</subject><subject>Genes</subject><subject>Genetic Engineering</subject><subject>Genetic modification</subject><subject>Genetically modified organisms</subject><subject>Genomes</subject><subject>House mouse</subject><subject>Mice</subject><subject>Mice, Inbred ICR</subject><subject>Microinjection</subject><subject>Microinjections</subject><subject>MicroRNA</subject><subject>MicroRNAs</subject><subject>MicroRNAs - genetics</subject><subject>MicroRNAs - metabolism</subject><subject>miRNA</subject><subject>Molecular Sequence Data</subject><subject>Oocytes</subject><subject>Oocytes - cytology</subject><subject>Oocytes - growth & development</subject><subject>Oocytes - metabolism</subject><subject>Plasmids</subject><subject>Poly(A)</subject><subject>Polyadenine</subject><subject>Polyadenylation</subject><subject>Pseudopregnancy</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>Rodents</subject><subject>Stem cells</subject><subject>Transcription activator-like effector nucleases</subject><subject>Xenopus</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNqNkl2L1DAUhoso7of-A9GCsODFjPlq09wIw7LqwLAL6-htSNPTToa2mU1Scf69Gae7TEFBcpFD8pw3J-e8SfIGozmmHH_c2sH1qp3vbA9zhHiOEHuWnGNBySwniD4_ic-SC--3CGW0yPOXyRlhmOUFEudJsVaugQBV2kAPaQUtBGP71NZpZ-5vFz41fYw0pOU-XS9WN7ep3_sA3avkRa1aD6_H_TL5_vlmff11trr7srxerGY6FyTMKoRFyXGBM460VrgQJSVIo5qhQitaA88IxprUGedEcAKCIo4oAqw04lzRy-TdUXfXWi_HT3uJGSMsz3mBI7E8EpVVW7lzplNuL60y8s-BdY1ULhjdghSVoFqwSlOILaC8ZAjXILTAQglBadT6NL42lB1UGvrgVDsRnd70ZiMb-1NSLuIHDsW8HwWcfRjAh3-UPFKNilWZvrZRTHfGa7lgvIhYxvJIzf9CxVVBnEgce23i-SThwyQhMgF-hUYN3svlt_v_Z-9-TNmrE3YDqg0bb9vh4BQ_BdkR1M5676B-6hxG8uDax27Ig2vl6NqY9va0609JjzalvwGTcuOI</recordid><startdate>20131016</startdate><enddate>20131016</enddate><creator>Takada, Shuji</creator><creator>Sato, Tempei</creator><creator>Ito, Yoshiaki</creator><creator>Yamashita, Satoshi</creator><creator>Kato, Tomoko</creator><creator>Kawasumi, Miyuri</creator><creator>Kanai-Azuma, Masami</creator><creator>Igarashi, Arisa</creator><creator>Kato, Tomomi</creator><creator>Tamano, Moe</creator><creator>Asahara, Hiroshi</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20131016</creationdate><title>Targeted gene deletion of miRNAs in mice by TALEN system</title><author>Takada, Shuji ; Sato, Tempei ; Ito, Yoshiaki ; Yamashita, Satoshi ; Kato, Tomoko ; Kawasumi, Miyuri ; Kanai-Azuma, Masami ; Igarashi, Arisa ; Kato, Tomomi ; Tamano, Moe ; Asahara, Hiroshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-d019b7181570cca189b320c0f408ca3fe75211c2f5772972e9307030e1ac077a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animal species</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Childrens health</topic><topic>Chromosomes</topic><topic>Clonal deletion</topic><topic>Deoxyribonucleases, Type II Site-Specific - genetics</topic><topic>Deoxyribonucleases, Type II Site-Specific - metabolism</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Intergenic - genetics</topic><topic>DNA, Intergenic - metabolism</topic><topic>DNA-Binding Proteins - genetics</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Embryo, Mammalian</topic><topic>Female</topic><topic>Gene Deletion</topic><topic>Gene mutation</topic><topic>Gene sequencing</topic><topic>Gene targeting</topic><topic>Gene Targeting - methods</topic><topic>Genes</topic><topic>Genetic Engineering</topic><topic>Genetic modification</topic><topic>Genetically modified organisms</topic><topic>Genomes</topic><topic>House mouse</topic><topic>Mice</topic><topic>Mice, Inbred ICR</topic><topic>Microinjection</topic><topic>Microinjections</topic><topic>MicroRNA</topic><topic>MicroRNAs</topic><topic>MicroRNAs - genetics</topic><topic>MicroRNAs - metabolism</topic><topic>miRNA</topic><topic>Molecular Sequence Data</topic><topic>Oocytes</topic><topic>Oocytes - 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Performed the experiments: ST TA YI SY TomokoK MK MK-A AI TomomiK MT. Analyzed the data: ST TA YI. Contributed reagents/materials/analysis tools: ST HA. Wrote the paper: ST HA.</notes><notes>Competing Interests: The authors have declared that no competing interests exist.</notes><abstract>Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the application of TALEN system for mice and successfully obtained gene targeting technique in mice for intergenic region and series of microRNAs. Microinjection of synthesized RNA of TALEN targeting each gene in one cell stage of embryo was carried out and injected oocytes were transferred into pseudopregnant ICR female mice, producing a high success rate of the targeted deletion of miRNA genes. In our condition, TALEN RNA without poly(A) tail worked better than that of with poly(A) tail. This mutated allele in miRNA was transmitted to the next generation, suggesting the successful germ line transmission of this targeting method. Consistent with our notion of miRNAs maturation mechanism, in homozygous mutant mice of miR-10a, the non- mutated strand of miRNAs expression was completely diminished. This method will lead us to expand and accelerate our genetic research using mice in a high throughput way.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24146809</pmid><doi>10.1371/journal.pone.0076004</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2013-10, Vol.8 (10), p.e76004 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1442466781 |
| source | Publicly Available Content Database; PubMed Central |
| subjects | Animal species Animals Base Sequence Childrens health Chromosomes Clonal deletion Deoxyribonucleases, Type II Site-Specific - genetics Deoxyribonucleases, Type II Site-Specific - metabolism Deoxyribonucleic acid DNA DNA, Intergenic - genetics DNA, Intergenic - metabolism DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism Embryo, Mammalian Female Gene Deletion Gene mutation Gene sequencing Gene targeting Gene Targeting - methods Genes Genetic Engineering Genetic modification Genetically modified organisms Genomes House mouse Mice Mice, Inbred ICR Microinjection Microinjections MicroRNA MicroRNAs MicroRNAs - genetics MicroRNAs - metabolism miRNA Molecular Sequence Data Oocytes Oocytes - cytology Oocytes - growth & development Oocytes - metabolism Plasmids Poly(A) Polyadenine Polyadenylation Pseudopregnancy Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Ribonucleic acid RNA Rodents Stem cells Transcription activator-like effector nucleases Xenopus |
| title | Targeted gene deletion of miRNAs in mice by TALEN system |
| url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-09-21T18%3A09%3A07IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Targeted%20gene%20deletion%20of%20miRNAs%20in%20mice%20by%20TALEN%20system&rft.jtitle=PloS%20one&rft.au=Takada,%20Shuji&rft.date=2013-10-16&rft.volume=8&rft.issue=10&rft.spage=e76004&rft.pages=e76004-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0076004&rft_dat=%3Cgale_plos_%3EA478246546%3C/gale_plos_%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c692t-d019b7181570cca189b320c0f408ca3fe75211c2f5772972e9307030e1ac077a3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=1442466781&rft_id=info:pmid/24146809&rft_galeid=A478246546&rfr_iscdi=true |