Overview
        The team aims to test subjects for the presence of metabolic syndrome-specific miRNAs at disease concentrations.Through DSN enzyme.circular DNA-RNA chimeric probe,DR]system and other steps.we can amplify the input of trace microRNA signals which are difficult to detect and convert them into easily detectable fluorescence signals,so as to determine whether the subjects have metabollc syndrome.At the same time,we hope to combine colloldal gold particles technology to achieve visual detection.On this page,we demonstrate the level at which our method is likely to work.
Can we achieve amplification of microRNA signals?
        If the microRNA target is present,it will hybridize to the probe and duplex-specific nuclease (DSN)will cleave DNA in DNA-RNA hybrid duplexes,re-releasing the microRNA to achieve linear signal amplification.At the same time,rest probe sequences (the RNA strand in the DNA-RNA molecule)will fold into sgRNA. After the completlon of the first reactlon,the reactlon llquld enters the second reaction.
Can we achieve signal transformation?
        The Input mlcroRNA slgnal Is converted Into sgRNA,whlch will form a ternary complex with Cas12a and asslstant DNA,activating Cas1 2a's trans cleavage ac- tivity.There are aggregated colloidal gold particles (aggregated through ssDNA connections)in the reaction solution,which can also be cleaved by activated Cas12a to release the visible signal.
Have we selected the right microRNAs that are specific of metab
        We screened three biomarkers of metabolic syndrome-related diseases respectively.
Literature searches were conducted in PubMed.EMBASE.Cochrane,Web of Science and Ovid from 1980 to April 2023.we performed the following search: 'micro RNA"OR "mIRNA"AND ("NAFLA"OR "non-alcohollc fatty liver disease")OR (DM"OR "dlabetes mellitus")OR (depression"OR "depressive disorder) AND "biomarker".

Our inclusion criteria:

(1).   

The study participants were patlents dlagnosed with NAFLD.DM,or depression.

(2).   

miRNAs are studied as diagnostic markers for diseases.

(3).   

Studies were performed in miRNAs detected in serum or plasma.

(4).   

The selected biomarkers should be easily detectable and detectable in the early stages of the disease,preferably at concentrations that change with the disease process.

        We first searched each database using keywords,removing duplicates that did not meet the inclusion criteria by reading the titles or abstracts.and then reading the articles in full to exclude studles whose results were not statistically significant.The final numbers of included studies related to the three diseas- es of NAFLD,DM,and depression were 9,13,and 5,respectively.
To screen for the most appropriate biomarkers,we made tables of the specifics of the miRNAs included in the study to facilitate comparison.The table in- cludes:
the names of the miRNAs
Roles in the metabolic syndrome
Region of the subject population
Biological material (plasma.serum)
Assay method (qRT-PCR)
Reference values or fold changes in concentration of miRNA as a biomarker in controls and patients
Testing difficulty level
Finally,three selected miRNAs were has-miR-34a in NAFLD,has-miR-155 in DM,and has-miR-1281 in depression.
In addition,we also performed a biogenic analysis to verify the correlation between mir34a and metabolic syndrome,demonstrating that mir34a can be used as a biomarker for MS.
        The database was searched for more information about the selected miRNAs,involving nucleotide numbers,target genes,functions,sequences,and their ex- pression in cell lines.The databases of choice include miRBase.TarBase,GeneCards.RefSeq.DIANA-LncBase.and RNAcentral.The results show that the three miRNAs we screened do not target human genes and meet the whitelisting requirements of iGEM.
How do we add colloidal gold particles technology?
        
        Colloidal gold particles solutions have their own color.We modify sulfhydryl groups at both ends of a single strand of DNA,and through gold-sulfur bonds. we connect colloidal gold particles together.changing the color of the solution.When the single strand DNA is cut by different amounts of activated cas12a. the color of the solutlon gradually changes.We can use the app we have developed to Identify different colors and Judge the health status of the subjects. At present,we have realized the cross-linking of colloidal gold particles and the development of app.
Can our technology be used to detect other diseases?
        
        Colloidal gold particles solutions have their own color.We modify sulfhydryl groups at both ends of a single strand of DNA,and through gold-sulfur bonds. we connect colloidal gold particles together.changing the color of the solution.When the single strand DNA is cut by different amounts of activated cas12a. the color of the solution gradually changes.We can use the app we have developed to identify different colors and judge the health status of the subjects. At present,we have reallzed the cross-linking of colloldal gold particles and the development of app.