earticle

영어

Gene silencing by targeting specific genes for degradation, particularly at the mRNA level, is an invaluable tool for gene function analysis and a powerful therapeutic strategy for human diseases, including cancer and viral infections. For degrading cytosolic RNAs is the use of
protein-based RNases and DNA/RNA-hydrolyzing mAbs, which can penetrate into living cells and degrade cytosolic RNAs. However, these approaches non sequence–specificity, leading to significant cytotoxicity. Based on a cell-penetrating, nucleic acid-hydrolyzing, 3D8 VL[1], we generated a synthetic library on the yeast surface by randomizing potential base interacting residues located in surface of c,c’,f-strands. And we considered that selected randomizing residues in framework by fixing several framework residues that affect directly or indirectly to CDR and antibody stability, like upper core and lower core, charged cluster. We isolated 3D8 VL variants with classical swine fever virus Npro and E2 genes –selective binding against 18-bp
single stranded (ss)-RNA substrates, selected 3D8 VL variants that had ~10-fold higher affinity and ~2-fold greater selective affinity for target ss-RNAs than for off targets.