A Novel Transgenic Mouse Line for Tracing MicroRNA-155-5p Activity In Vivo
MicroRNA-155 (miR-155) plays significant role in various physiological processes involving both innate and adaptive immunity. miR-155 expression level changes dynamically during various immune responses. However, current approaches for miR-155 detection at the RNA level do not precisely reflect the...
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| Veröffentlicht in: | PloS one 2015-06, Vol.10 (6), p.e0128198-e0128198 |
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| creator | Phiwpan, Krung Guo, Jie Zhang, Wei Hu, Tanyu Boruah, Bhargavi M Zhang, Jianhua Zhou, Xuyu |
| description | MicroRNA-155 (miR-155) plays significant role in various physiological processes involving both innate and adaptive immunity. miR-155 expression level changes dynamically during various immune responses. However, current approaches for miR-155 detection at the RNA level do not precisely reflect the real-time activity. Herein, we generated a transgenic mouse line (R26-DTR-155T) for determination of miR-155-5p activity in vivo by inserting miR-155-5p target sequence downstream of a reporter transgene comprising Diphtheria Toxin Receptor and TagBlue fluorescence protein. Using this approach, R26-DTR-155T mice were able to measure variation in levels of miR-155-5p activity in specific cell types of interest. The DTR expression levels were inversely correlated with the endogenous miR-155 expression pattern as detected by quantitative RT-PCR. Our data demonstrate a novel transgenic mouse line which could be useful for tracing miR-155-5p activity in specific cell types through measurement of miR-155-5p activity at single cell level. |
| doi_str_mv | 10.1371/journal.pone.0128198 |
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However, current approaches for miR-155 detection at the RNA level do not precisely reflect the real-time activity. Herein, we generated a transgenic mouse line (R26-DTR-155T) for determination of miR-155-5p activity in vivo by inserting miR-155-5p target sequence downstream of a reporter transgene comprising Diphtheria Toxin Receptor and TagBlue fluorescence protein. Using this approach, R26-DTR-155T mice were able to measure variation in levels of miR-155-5p activity in specific cell types of interest. The DTR expression levels were inversely correlated with the endogenous miR-155 expression pattern as detected by quantitative RT-PCR. Our data demonstrate a novel transgenic mouse line which could be useful for tracing miR-155-5p activity in specific cell types through measurement of miR-155-5p activity at single cell level.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0128198</identifier><identifier>PMID: 26030404</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>3' Untranslated Regions - genetics ; Adaptive immunity ; Animals ; Antigens ; Artificial chromosomes ; B cells ; Brain research ; Cell Lineage ; Chromosomes, Artificial, Bacterial - genetics ; Cloning ; Diphtheria ; Diphtheria toxin ; Diphtheria Toxin - genetics ; Fluorescence ; Gene expression ; Genes, Reporter - genetics ; Genetic engineering ; Genetic Engineering - methods ; Immune response ; Immunity ; Immunology ; In vivo methods and tests ; Laboratories ; Luminescent Proteins - genetics ; Lymphocytes ; Mice ; Mice, Transgenic ; MicroRNA ; MicroRNAs ; MicroRNAs - genetics ; miRNA ; Physiological aspects ; Polymerase chain reaction ; Proteins ; Ribonucleic acid ; RNA ; RNA polymerase ; Rodents ; Sensors ; Transgenic animals ; Transgenic mice</subject><ispartof>PloS one, 2015-06, Vol.10 (6), p.e0128198-e0128198</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Phiwpan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://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>2015 Phiwpan et al 2015 Phiwpan et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-5bf86bb299dcc19f22b7f1699d23c5a168c36414391274ea3d130ce0f4cd7edf3</citedby><cites>FETCH-LOGICAL-c692t-5bf86bb299dcc19f22b7f1699d23c5a168c36414391274ea3d130ce0f4cd7edf3</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/PMC4452368/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4452368/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2100,2919,23857,27915,27916,53782,53784,79361,79362</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26030404$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Ahuja, Sunil K.</contributor><creatorcontrib>Phiwpan, Krung</creatorcontrib><creatorcontrib>Guo, Jie</creatorcontrib><creatorcontrib>Zhang, Wei</creatorcontrib><creatorcontrib>Hu, Tanyu</creatorcontrib><creatorcontrib>Boruah, Bhargavi M</creatorcontrib><creatorcontrib>Zhang, Jianhua</creatorcontrib><creatorcontrib>Zhou, Xuyu</creatorcontrib><title>A Novel Transgenic Mouse Line for Tracing MicroRNA-155-5p Activity In Vivo</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>MicroRNA-155 (miR-155) plays significant role in various physiological processes involving both innate and adaptive immunity. miR-155 expression level changes dynamically during various immune responses. However, current approaches for miR-155 detection at the RNA level do not precisely reflect the real-time activity. Herein, we generated a transgenic mouse line (R26-DTR-155T) for determination of miR-155-5p activity in vivo by inserting miR-155-5p target sequence downstream of a reporter transgene comprising Diphtheria Toxin Receptor and TagBlue fluorescence protein. Using this approach, R26-DTR-155T mice were able to measure variation in levels of miR-155-5p activity in specific cell types of interest. The DTR expression levels were inversely correlated with the endogenous miR-155 expression pattern as detected by quantitative RT-PCR. Our data demonstrate a novel transgenic mouse line which could be useful for tracing miR-155-5p activity in specific cell types through measurement of miR-155-5p activity at single cell level.</description><subject>3' Untranslated Regions - genetics</subject><subject>Adaptive immunity</subject><subject>Animals</subject><subject>Antigens</subject><subject>Artificial chromosomes</subject><subject>B cells</subject><subject>Brain research</subject><subject>Cell Lineage</subject><subject>Chromosomes, Artificial, Bacterial - genetics</subject><subject>Cloning</subject><subject>Diphtheria</subject><subject>Diphtheria toxin</subject><subject>Diphtheria Toxin - genetics</subject><subject>Fluorescence</subject><subject>Gene expression</subject><subject>Genes, Reporter - genetics</subject><subject>Genetic engineering</subject><subject>Genetic Engineering - methods</subject><subject>Immune response</subject><subject>Immunity</subject><subject>Immunology</subject><subject>In vivo methods and tests</subject><subject>Laboratories</subject><subject>Luminescent Proteins - genetics</subject><subject>Lymphocytes</subject><subject>Mice</subject><subject>Mice, Transgenic</subject><subject>MicroRNA</subject><subject>MicroRNAs</subject><subject>MicroRNAs - genetics</subject><subject>miRNA</subject><subject>Physiological aspects</subject><subject>Polymerase chain reaction</subject><subject>Proteins</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA polymerase</subject><subject>Rodents</subject><subject>Sensors</subject><subject>Transgenic animals</subject><subject>Transgenic mice</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNkl1v0zAYhSMEYmPwDxBEQkJwkeLvxDdI1TSgqNukMXZrOY6dukrtLk6q7d_j0Gxq0C6QL_z1nGP79UmStxDMIM7hl7XvWyeb2dY7PQMQFZAXz5JjyDHKGAL4-cH4KHkVwhoAigvGXiZHiAEMCCDHyc95euF3ukmvW-lCrZ1V6bnvg06X1unU-HbYUdbV6blVrb-6mGeQ0oxu07nq7M529-nCpTd2518nL4xsgn4z9ifJ729n16c_suXl98XpfJkpxlGX0dIUrCwR55VSkBuEytxAFqcIKyohKxRmBBLMIcqJlriCGCgNDFFVriuDT5L3e99t44MYyxBEFBLOcQ7ySCz2ROXlWmxbu5HtvfDSir8Lvq2FbDurGi04BVrykjEEC1JSIzXHppIYI81JoQevr-NpfbnRldKua2UzMZ3uOLsStd8JQijCrIgGn0aD1t_2OnRiY4PSTSOdjoUe7k1zyginEf3wD_r060aqlvEB1hkfz1WDqZgThAtMKUORmj1BxVbpjVUxM8bG9Yng80QQmU7fdbXsQxCLX1f_z17eTNmPB-xKy6ZbBd_0nfUuTEGyB2PMQmi1eSwyBGKI_EM1xBB5MUY-yt4dftCj6CHj-A94-PiL</recordid><startdate>20150601</startdate><enddate>20150601</enddate><creator>Phiwpan, Krung</creator><creator>Guo, Jie</creator><creator>Zhang, Wei</creator><creator>Hu, Tanyu</creator><creator>Boruah, Bhargavi M</creator><creator>Zhang, Jianhua</creator><creator>Zhou, Xuyu</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>AEUYN</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>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20150601</creationdate><title>A Novel Transgenic Mouse Line for Tracing MicroRNA-155-5p Activity In Vivo</title><author>Phiwpan, Krung ; Guo, Jie ; Zhang, Wei ; Hu, Tanyu ; Boruah, Bhargavi M ; Zhang, Jianhua ; Zhou, Xuyu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-5bf86bb299dcc19f22b7f1699d23c5a168c36414391274ea3d130ce0f4cd7edf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>3' Untranslated Regions - genetics</topic><topic>Adaptive immunity</topic><topic>Animals</topic><topic>Antigens</topic><topic>Artificial chromosomes</topic><topic>B cells</topic><topic>Brain research</topic><topic>Cell Lineage</topic><topic>Chromosomes, Artificial, Bacterial - genetics</topic><topic>Cloning</topic><topic>Diphtheria</topic><topic>Diphtheria toxin</topic><topic>Diphtheria Toxin - genetics</topic><topic>Fluorescence</topic><topic>Gene expression</topic><topic>Genes, Reporter - genetics</topic><topic>Genetic engineering</topic><topic>Genetic Engineering - methods</topic><topic>Immune response</topic><topic>Immunity</topic><topic>Immunology</topic><topic>In vivo methods and tests</topic><topic>Laboratories</topic><topic>Luminescent Proteins - genetics</topic><topic>Lymphocytes</topic><topic>Mice</topic><topic>Mice, Transgenic</topic><topic>MicroRNA</topic><topic>MicroRNAs</topic><topic>MicroRNAs - genetics</topic><topic>miRNA</topic><topic>Physiological aspects</topic><topic>Polymerase chain reaction</topic><topic>Proteins</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>RNA polymerase</topic><topic>Rodents</topic><topic>Sensors</topic><topic>Transgenic animals</topic><topic>Transgenic mice</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Phiwpan, Krung</creatorcontrib><creatorcontrib>Guo, Jie</creatorcontrib><creatorcontrib>Zhang, Wei</creatorcontrib><creatorcontrib>Hu, Tanyu</creatorcontrib><creatorcontrib>Boruah, Bhargavi M</creatorcontrib><creatorcontrib>Zhang, Jianhua</creatorcontrib><creatorcontrib>Zhou, Xuyu</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology 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>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection (ProQuest)</collection><collection>Natural Science Collection (ProQuest)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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However, current approaches for miR-155 detection at the RNA level do not precisely reflect the real-time activity. Herein, we generated a transgenic mouse line (R26-DTR-155T) for determination of miR-155-5p activity in vivo by inserting miR-155-5p target sequence downstream of a reporter transgene comprising Diphtheria Toxin Receptor and TagBlue fluorescence protein. Using this approach, R26-DTR-155T mice were able to measure variation in levels of miR-155-5p activity in specific cell types of interest. The DTR expression levels were inversely correlated with the endogenous miR-155 expression pattern as detected by quantitative RT-PCR. Our data demonstrate a novel transgenic mouse line which could be useful for tracing miR-155-5p activity in specific cell types through measurement of miR-155-5p activity at single cell level.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>26030404</pmid><doi>10.1371/journal.pone.0128198</doi><oa>free_for_read</oa></addata></record> |
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| subjects | 3' Untranslated Regions - genetics Adaptive immunity Animals Antigens Artificial chromosomes B cells Brain research Cell Lineage Chromosomes, Artificial, Bacterial - genetics Cloning Diphtheria Diphtheria toxin Diphtheria Toxin - genetics Fluorescence Gene expression Genes, Reporter - genetics Genetic engineering Genetic Engineering - methods Immune response Immunity Immunology In vivo methods and tests Laboratories Luminescent Proteins - genetics Lymphocytes Mice Mice, Transgenic MicroRNA MicroRNAs MicroRNAs - genetics miRNA Physiological aspects Polymerase chain reaction Proteins Ribonucleic acid RNA RNA polymerase Rodents Sensors Transgenic animals Transgenic mice |
| title | A Novel Transgenic Mouse Line for Tracing MicroRNA-155-5p Activity In Vivo |
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