Screening And Identification Of Age-related CircRNA In The Human Whole Blood

Research background:Forensic age estimation is of great significance in forensic case detection.With the development of molecular biology,a series of aging-related biomarkers was discovered by researchers as candidate markers for forensic age estimation.However,age estimation based on these markers has some problems,such as poor accuracy,poor repeatability,and limited object of application,which result in the limitation of forensic application value.Individual aging is a complex biological process.It is not easy to use a single age-inference marker to carry out accurate age estimation.We need to use the big-data platform to introduce multiple age-inference markers to implement joint multivariate analysis.Therefore,it is still very important to continue to look for a stable age predictor with an obvious trend of age that have a wide range of application.Recent studies have shown that circ RNA,which is more stable than m RNA,is widely expressed in a variety of tissues of the human body and is involved in regulating the aging process of the body.The expression of circ RNA may become a new age inference marker,and its expression is related with age.Based on this,the screening and identification of age-related circ RNA markers may become an important basic research content of forensic age estimation.Objectives:To screen and identify obvious age-related circ RNA in the whole transcriptome to provide candidate biomarkers for forensic age estimation.Methods:?1?Fresh blood samples were collected from relatively healthy people from different ages.?2?The circ RNA expression profiles of whole blood of young,middle-aged and elderly groups were detected by using circ RNA microarray,and then the significantly age-related circ RNA markers were analyzed and screened using bioinformatics methods.?3?The circ RNA sequences were obtained from circ Base website or the circ RNA data provided by the supplier of circ RNA microarray,and the expression profile of every selected circ RNA marker was verified by the RT-q PCR method.Results:?1?According to the results of circ RNA microarray,three age-related expression profiles,which were named as Profile 14,Profile 15 and Profile 9 separately,were screened with the combination of group-comparison analysis and STC.Finally,5,10 and 10 circ RNA candidates were selected for later validation of Profile 14,Profile 15 and Profile 9 respectively.?2?Among the 25 selected candidate circ RNAs,17 circ RNAs were successfully detected by q PCR,and the overall expression profile with aging of q PCR data were consistent with the results of circ RNA microarray.The expression of circ RNA hsa_circ₀003416,hsa_circ₀00685 and hsa_circ₀140253 decrease gradually with aging,which meets the screening requirements for biological age estimation.Conclusion:Some target circ RNAs obtained from the whole transcriptome of human blood show age-related expression and could be used as candidate biomarkers for forensic age estimation.