MOFzyme Of Payload G4:in Vitro Real-time And Ultra-sensitive Chemiluminescence With Dual Amplification Function For The Detection Of Acute Myocardial Infarction Related MiRNAs

Micro RNA(miRNA)is one kind of small,non-coding RNA,which is about 18-25 nucleotides in length.In animals,the vast majority of miRNAs can combine with the complementary area of the target genes in 3'-UTR(non-coding regions)to suppress the conversion from target genes to proteins.In this way,miRNAs can affect the growth and development of organisms in the level of cells,tissues or even individuals,as well as participate in a variety of disease processes at the same time.Thus,the detection of disease related miRNAs can play a constructive role in cancer researches.However,the amount of miRNAs is very small,and it is usually difficult to detect the disease related miRNAs directly in real samples.Therefore,this paper will mainly focus on solving these problems,establishing a chemiluminescence platform for the ultra-sensitive and real-time detection of acute myocardial infarction related miRNAs.The whole research can be roughly divided into the following two parts:The first part is the construction of G4/MOFzyme CL detection platform.We mainly use hemin as the organic ligand and zinc as the metal ligand to synthesize the hemin bridged metal-organic framework(MOF)material.Due to the presence of hemin,this newly synthesized MOF material can provide copious amount of binding sites for G-quadruplexes(G4s),in order for the payload of G4/hemin DNAzyme on the surface of MOF.On this basis,the synthesized Zn-hemin MOF material can be combined with rolling circle amplification(RCA)strategy,that is using the target miRNAs(miRNA-133a,miRNA-499)as primer to amplify the well-designed circular template in the presence of dNTPs and phi29 DNA polymerase,generating a series of G4 polymers.These G4 polymers can bind to hemin which locates on the surface of MOF material to form G4/MOFzyme.The second part is the application of G4/MOFzyme,it can boost the CL of Luminol-H2O2,and generate extended CL which exceeds 24 hours.At the same time,G4/MOFzyme catalysed CL intensity is directly proportional to the concentration of the target miRNAs,and the results can be seen even with naked eyes and captured by a smartphone.The detection limit can be as low as 1fM with cellphone imaging and 2aM with CL instrument,which is much more superior than most of the methods reported in literatures.Besides,the constructed CL sensing platform also has good specificity,it can distinguish between complementary and noncomplementary miRNAs.Furthermore,it has a strong anti-interference ability,which obtains good detection results in 10%human serum.In summary,the establishment of this G4/MOFzyme chemiluminescence sensing platform can provide a new analytical method for the detection of miRNAs,and meantime has important application potential in the diagnosis of acute myocardial infarction.