法医器官组织特异性circRNAs的筛选及确证

Screening and confirmation of circRNA markers for forensic organ tissue identification

目的筛选并确证器官组织特异性circRNAs分子标记物,应用于法医器官组织溯源。方法从TSCD数据库及文献中,筛选出器官组织特异性circRNAs,设计并验证引物。收集心脏、大脑、肝脏、皮肤、骨骼肌、肺脏等人体组织样本,经总RNA提取、RT-qPCR、琼脂糖凝胶电泳等步骤对候选circRNAs进行组织表达水平分析,并评估方法的灵敏度。结果利用数据库和文献初步筛选出25个器官组织特异性circRNAs。经验证,其中8个circRNAs在目标组织中的相对表达水平显著高于其他组织(P<0.01)。RT-qPCR方法在最低0.1ng起始RNA模板量情况下可在所有目标组织检出特异性circRNAs。结论circRNAs可以作为鉴识法医器官组织新的分子标记,值得继续深入研究。

Objective To screen and confirm organ tissue specific circRNA markers for forensic organ tissue identification.Methods Organ tissue specific circRNAs were screened from TSCD database and literature.Primers were designed and then verified.Total RNA was extracted from human organ tissue samples such as heart,brain,liver,skin,skeletal muscle and lung,and the expression levels of candidate circRNAs were analyzed by RT-qPCR and agarose gel electrophoresis.Analytical sensitivity of the assays was evaluated.Results Twenty-five potential specific circRNAs were selected from TSCD database and literature,eight of which were confirmed over-expressed in target tissues(P<0.01).Using the RT-qPCR assay,all of the confirmed markers were detectable in the target organ tissue when the amount of RNA was as low as 0.1 ng.Conclusion CircRNA can be used as a new promising biomarker for organ tissue identification in forensic medicine.