{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T11:58:21Z","timestamp":1783079901230,"version":"3.54.6"},"reference-count":47,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,11,26]],"date-time":"2024-11-26T00:00:00Z","timestamp":1732579200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"BB21 Plus"},{"name":"Busan Metropolitan City"},{"name":"Busan Techno Park"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["ncRNA"],"abstract":"<jats:p>Background: When a body is discovered at a crime or murder scene, it is crucial to examine the body and estimate its postmortem interval (PMI). Accurate estimation of PMI is vital for identifying suspects and providing clues to resolve the case. MicroRNAs (miRNAs or miRs) are small non-coding RNAs that remain relatively stable in the cell nucleus even after death-related changes occur. Objective: This study developed a molecular beacon probe for mmu-miR-133a-5p and assessed its use in mouse muscle tissue at temperatures of 4 \u00b0C and 21 \u00b0C to estimate the PMI. Methods: A total of 36 healthy adult male BALB\/c mice were divided into 9 PMI time points (0, 2, 6, 8, and 10 days) with 3 mice per time point, and they were exposed to 4 \u00b0C and 21 \u00b0C. Next, the expression pattern of mmu-miR-133a in the skeletal muscle tissue over a 10-day PMI period was analyzed using the developed molecular beacon probe. Results: The molecular beacon (MB) probe was designed for optimal thermodynamic stability with a hairpin structure that opened in the presence of mmu-miR-133a-5p, thus separating the fluorophore from the quencher and resulting in a strong fluorescence signal at 495 nm. Fluorescence intensity increased with mmu-miR-133a-5p concentration from 1 ng\/\u03bcL to 1000 ng\/\u03bcL and exhibited a strong correlation (R2 = 0.9966) and a detection limit of 1 ng\/\u03bcL. Subsequently, the expression level of mmu-miR-133a-5p was observed to be stable in mouse skeletal muscle tissue at both 4 \u00b0C and 21 \u00b0C. Conclusions: This user-friendly assay can complete measurements in just 30 min after RNA extraction and is suitable for point-of-care testing, and it possesses the potential to improve existing complex and time-consuming methods for PMI estimation.<\/jats:p>","DOI":"10.3390\/ncrna10060058","type":"journal-article","created":{"date-parts":[[2024,11,26]],"date-time":"2024-11-26T04:02:18Z","timestamp":1732593738000},"page":"58","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Detection of miR-133a-5p Using a Molecular Beacon Probe for Investigating Postmortem Intervals"],"prefix":"10.3390","volume":"10","author":[{"given":"Eun Hye","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingyoung","family":"Jeong","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Project, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4022-3415","authenticated-orcid":false,"given":"Kwangmin","family":"Park","sequence":"additional","affiliation":[{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Project, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong Geon","family":"Lee","sequence":"additional","affiliation":[{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Project, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eun Ju","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Korea Mycobacterium Resource Center (KMRC), Department of Research and Development, The Korean Institute of Tuberculosis, Osong 28158, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haneul","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Project, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ah Yeoung","family":"Kim","sequence":"additional","affiliation":[{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Project, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jae Won","family":"Ahn","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5790-5746","authenticated-orcid":false,"given":"Hyun Jun","family":"Woo","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science, Semyung University, Jecheon 27136, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2511-6555","authenticated-orcid":false,"given":"Sunghyun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Project, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6905-5710","authenticated-orcid":false,"given":"Jaewon","family":"Lim","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Project, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jungho","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Project, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1007\/s004149900090","article-title":"Experiences with a compound method for estimating the time since death. 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