The increased potential for gene doping since the introduction of gene therapy presents the need to develop antidoping assays. We therefore aimed to develop a quick and simple method for the detection of specifically targeted exogenous doping genes utilizing an in vitro clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9 (CRISPR-Cas9) system. A human erythropoietin (hEPO) is a drug frequently used for doping in athletes, and gene doping using gene transfer techniques may be attempted. Therefore, we selected hEPO gene as a model of exogenous doping gene, and complemental single guide RNA (sgRNA) was designed to specifically bind to the four exon-exon junctions in the hEPO cDNA. For the rapid reaction of CRISPR-Cas9, further optimization was performed using an open-source program (CRISPOR) that avoids TT and GCC motifs before the protospacer adjacent motif (PAM) domain and predicts the efficiency of the sgRNA. We optimized the in vitro Cas9 assay and dual use of sgRNA for double cleavage and identified the limit of detection (LOD) of the 1.25 nM of the double cleavage method. We expect that the improved CRISPR-Cas9 method can be used for antidoping analysis of gene doping.
This study aimed to produce cellulose-based conductive fabrics with electrical conductivity and flexibility. Bacterial cellulose (BC) and three chemical cellulose (CC), namely methyl cellulose (MC), hydroxypropyl cellulose (HPMC) and carboxymethyl cellulose (CMC) were in situ polymerized with aniline and the four conductive cellulose fabrics were compared and evaluated. Matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy analysis confirmed that three CC-PANI composites displayed longer and more stable polymerization pattern than BC-PANI because of the different polymerization method: bulk polymerization for BC-PANI and emulsion polymerization for CC-PANI, respectively. The electrical conductivity of BC-PANI and CC-PANI were ranging from 0.962 × 10 −2 S/cm to 2.840 × 10 −2 S/cm. MC-PANI showed the highest electrical conductivity among the four conductive cellulose fabrics. The flexibility and crease recovery results showed that MC-PANI had the highest flexibility compared to BC-PANI, HPMC-PANI, and CMC-PANI. These results have confirmed that the electrical conductivity and flexibility were influenced by the type of cellulose, and MC-PANI was found to have the best performance in the electrical conductivity and flexibility.
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.