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miR-26a/30d/152 are reliable reference genes for miRNA quantification in skin wound age estimation 认领 引用
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作者 Longlong Suo Jian Cheng +10 位作者 Haomiao Yuan Zhenfei Jiang Dilichati Tash Linlin Wang Hao Cheng Zhongduo Zhang Fuyuan Zhang Miao Zhang Zhipeng Cao Rui Zhao Dawei Guan 《Forensic Sciences Research》 CSCD 2023年第3期230-240,共11页
MicroRNAs(miRNAs)are a class of small non-coding RNAs that exert their biological functions as negative regulators of gene expression.They are involved in the skin wound healing process with a dynamic expression patte... MicroRNAs(miRNAs)are a class of small non-coding RNAs that exert their biological functions as negative regulators of gene expression.They are involved in the skin wound healing process with a dynamic expression pattern and can therefore potentially serve as biomarkers for skin wound age estimation.However,no reports have described any miRNAs as suitable reference genes(RGs)for miRNA quantification in wounded skin or samples with post-mortem changes.Here,we aimed to identify specific miRNAs as RGs for miRNA quantification to support further studies of skin wound age estimation.Overall,nine miRNAs stably expressed in mouse skin at certain posttraumatic intervals(PTls)were preselected by next-generation sequencing as candidate RGs.These nine miRNAs and the commonly used reference genes(comRGs:U6,GAPDH,ACTB,18S,5S,LC-Ogdh)were quantitatively examined using quantitative real-time reverse-transcription polymerase chain reaction at different PTls during skin wound healing in mice.The stabilities of these genes were evaluated using four independent algorithms:GeNorm,NormFinder,BestKeeper,and comparative Delta Ct.Stability was further evaluated in mice with different post-mortem intervals(PMls).Overall,mmu-miR-26a-5p,mmu-miR-30d-5p,and mmu-miR-152-3p were identified as the most stable genes at both different PTIs and PMls.These three miRNA RGs were additionally validated and compared with the comRGs in human samples.After assessing using one,two,or three miRNAs in combination for stability at different PTls,PMls,or in human samples,the set of miR-26a/30d/152 was approved as the best normalizer.In conclusion,our data suggest that the combination of miR-26a/30d/152 is recommended as the normalization strategy for miRNA qRT-PCR quantification in skin wound age estimation. 展开更多
关键词 forensic sciences forensic genetics reference gene miRNA wound age estimation skin wound healing normalization qRT-PCR
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