Aptamers have attracted much attention as the next generation of affinity reagents. Unfortunately, the selection efficiency remains a critical bottleneck for the widespread application of aptamers. Herein, to accelerate aptamers discovery, a multifunctional microfluidic selection platform was developed, on which the selection efficiency was greatly improved and high-affinity and -specificity aptamers were generated within two round selections. The multifunctional screening platform, precisely manipulating magnetic beads on the micrometer scale, improved selection performance based on microfluidic continuous flow and enhanced the selection process control via in situ monitoring and real-time evaluation. This method could suppress ∼50-fold nonspecific binding nucleic acids compared to the conventional methods, further eliminate weakly bound nucleic acids within 9 min, and simultaneously perform the negative selection and positive selection. And the selection effectiveness was in situ and real-time monitoring. Three aptamers showed high affinity and specificity toward mucin 1 (MUC1) with dissociation constants (K) in nanomolar range (from 22 to 65 nM). Furthermore, the selected aptamer was able to specially label cancer cells and efficiently capture exosomes with 64% capture efficiency. It demonstrated that the multifunctional screening platform was an efficient method to generate high-quality aptamers in a rapid and economic manner.
BackgroundInterferon Regulatory Factor 6 (IRF6) gene encodes a member of the IRF family of transcription factors. Mutations in IRF6 cause Van der Woude Syndrome (VWS), which is the most common malformation of syndromic orofacial clefts in humans.MethodsHere, we performed sequencing studies of six families with VWS in the Chinese Han population. The entire IRF6‐coding region and the exon–intron boundaries including exons 3–8 and part of exon 9 were screened among all the collected family members by Sanger sequencing.ResultsWe found a novel splice site variant c.175‐6T>A, two novel missense variants (p.Lys66Arg and p.Pro107Thr), in addition with a previously reported missense variant (p.Leu87Phe), which were all located in and nearby exon 4 of IRF6. Meanwhile, a novel frameshift variant p.G257Vfs*46 in exon 7 of IRF6 was also detected. All the mutations presented to be co‐segregated in each family.ConclusionOur study has advanced the understanding of the genetic architecture of VWS and provides the basis for genetic counseling, antenatal diagnosis, and gene therapy of high risk groups.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.