{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T13:24:29Z","timestamp":1789997069988,"version":"4.0.1"},"reference-count":25,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"2","content-domain":{"domain":["www.ahajournals.org"],"crossmark-restriction":true},"short-container-title":["ATVB"],"published-print":{"date-parts":[[2015,2]]},"abstract":"<jats:sec>\n            <jats:title>Objective\u2014<\/jats:title>\n            <jats:p>\n              To ascertain the importance of a single regulatory element in the control of\n              <jats:italic>Cnn1<\/jats:italic>\n              expression using CRISPR\/Cas9 (clustered regularly interspaced short palindromic repeats\/CRISPR-associated protein 9) genome editing.\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Approach and Results\u2014<\/jats:title>\n            <jats:p>\n              The CRISPR\/Cas9 system was used to produce 3 of 18 founder mice carrying point mutations in an intronic CArG box of the smooth muscle cell\u2013restricted\n              <jats:italic>Cnn1<\/jats:italic>\n              gene. Each founder was bred for germline transmission of the mutant CArG box and littermate interbreeding to generate homozygous mutant (\n              <jats:italic>Cnn1<\/jats:italic>\n              <jats:sup>\u0394CArG\/\u0394CArG<\/jats:sup>\n              ) mice. Quantitative reverse transcription polymerase chain reaction, Western blotting, and confocal immunofluorescence microscopy showed dramatic reductions in\n              <jats:italic>Cnn1<\/jats:italic>\n              mRNA and CNN1 protein expression in\n              <jats:italic>Cnn1<\/jats:italic>\n              <jats:sup>\u0394CArG\/\u0394CArG<\/jats:sup>\n              mice with no change in other smooth muscle cell\u2013restricted genes and little evidence of off-target edits elsewhere in the genome. In vivo chromatin immunoprecipitation assay revealed a sharp decrease in binding of serum response factor to the mutant CArG box. Loss of CNN1 expression was coincident with an increase in Ki-67 positive cells in the normal vessel wall.\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions\u2014<\/jats:title>\n            <jats:p>\n              CRISPR\/Cas9 genome editing of a single CArG box nearly abolishes\n              <jats:italic>Cnn1<\/jats:italic>\n              expression in vivo and evokes increases in smooth muscle cell DNA synthesis. This facile genome editing system paves the way for a new generation of studies designed to test the importance of individual regulatory elements in living animals, including regulatory variants in conserved sequence blocks linked to human disease.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1161\/atvbaha.114.305017","type":"journal-article","created":{"date-parts":[[2014,12,24]],"date-time":"2014-12-24T05:19:40Z","timestamp":1419398380000},"page":"312-315","update-policy":"https:\/\/doi.org\/10.1161\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":49,"title":["CRISPR-Cas9 Genome Editing of a Single Regulatory Element Nearly Abolishes Target Gene Expression in Mice\u2014Brief Report"],"prefix":"10.1161","volume":"35","author":[{"given":"Yu","family":"Han","sequence":"first","affiliation":[{"name":"From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, Rochester, NY (Y.H., O.J.S., C.K.C., J.M.M.); and Jackson Laboratories, Bar Harbor, ME (A.W.C.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Orazio J.","family":"Slivano","sequence":"additional","affiliation":[{"name":"From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, Rochester, NY (Y.H., O.J.S., C.K.C., J.M.M.); and Jackson Laboratories, Bar Harbor, ME (A.W.C.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine K.","family":"Christie","sequence":"additional","affiliation":[{"name":"From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, Rochester, NY (Y.H., O.J.S., C.K.C., J.M.M.); and Jackson Laboratories, Bar Harbor, ME (A.W.C.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Albert W.","family":"Cheng","sequence":"additional","affiliation":[{"name":"From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, Rochester, NY (Y.H., O.J.S., C.K.C., J.M.M.); and Jackson Laboratories, Bar Harbor, ME (A.W.C.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph M.","family":"Miano","sequence":"additional","affiliation":[{"name":"From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, Rochester, NY (Y.H., O.J.S., C.K.C., J.M.M.); and Jackson Laboratories, Bar Harbor, ME (A.W.C.)."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.ge.20.120186.002341"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1008918913249"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1138140"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2501"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/cr.2014.11"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2652"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2013.04.025"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1093\/jmcb\/mjt047"},{"key":"e_1_3_5_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.01.027"},{"key":"e_1_3_5_11_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1231143"},{"key":"e_1_3_5_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2013.08.022"},{"key":"e_1_3_5_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2842"},{"key":"e_1_3_5_14_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1222794"},{"key":"e_1_3_5_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2828(03)00110-X"},{"key":"e_1_3_5_16_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.275.13.9814"},{"key":"e_1_3_5_17_2","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.111.232785"},{"key":"e_1_3_5_18_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1349-7006.2005.00118.x"},{"key":"e_1_3_5_19_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0508566103"},{"key":"e_1_3_5_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrg1619"},{"key":"e_1_3_5_21_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.111.262717"},{"key":"e_1_3_5_22_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1228606"},{"key":"e_1_3_5_23_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1241006"},{"key":"e_1_3_5_24_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.510362"},{"key":"e_1_3_5_25_2","doi-asserted-by":"publisher","DOI":"10.1096\/fj.09-140749"},{"key":"e_1_3_5_26_2","doi-asserted-by":"publisher","DOI":"10.1152\/physiolgenomics.00098.2011"}],"container-title":["Arteriosclerosis, Thrombosis, and Vascular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ahajournals.org\/doi\/full\/10.1161\/ATVBAHA.114.305017","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,12]],"date-time":"2024-05-12T23:18:06Z","timestamp":1715555886000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ahajournals.org\/doi\/10.1161\/ATVBAHA.114.305017"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,2]]},"references-count":25,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2015,2]]}},"alternative-id":["10.1161\/ATVBAHA.114.305017"],"URL":"https:\/\/doi.org\/10.1161\/atvbaha.114.305017","relation":{},"ISSN":["1079-5642","1524-4636"],"issn-type":[{"value":"1079-5642","type":"print"},{"value":"1524-4636","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,2]]},"assertion":[{"value":"2014-11-20","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-12-11","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-12-23","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}