In vitro selection of novel RNA ligands that bind human cytomegalovirus and block viral infection.
Ribonuclease-resistant RNA molecules that bind to infectious human cytomegalovirus (HCMV) were isolated in vitro from a pool of randomized sequences after 16 cycles of selection and amplification. The two ligands (L13 and L19) characterized exhibited high HCMV-binding affinity in vitro and effectively inhibited viral infection in tissue culture. Their antiviral activity was also specific as they only reacted with two different strains of HCMV but not with the related herpes simplex virus 1 and human cells. These two ligands appeared to function as antivirals by blocking viral entry. Ultraviolet (UV) crosslinking studies suggested that L13 and L19 bind to HCMV essential glycoproteins B and H, respectively. Thus, RNA ligands that bind to different surface antigens of HCMV can be simultaneously isolated by the selection procedure. Our study demonstrates the feasibility of using these RNA ligands as a research tool to identify viral proteins required for infectivity and as an antiviral agent to block viral infection.
Wang
J
J
Program in Infectious Diseases and Immunity, University of California, Berkeley 94720, USA.
Jiang
H
H
Liu
F
F
D000998:Antiviral Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism* / Q000472:pathogenicity; D004279:DNA, Viral / Q000096:biosynthesis; D019020:Directed Molecular Evolution; D018259:Herpesvirus 1, Human / Q000187:drug effects; D006801:Humans; D008024:Ligands; D011485:Protein Binding; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D012260:Ribonucleases / Q000378:metabolism; D017423:Sequence Analysis, RNA; D013379:Substrate Specificity; D013816:Thermodynamics; D014759:Viral Envelope Proteins / Q000378:metabolism; D010948:Viral Plaque Assay
D000998:Antiviral Agents; D004279:DNA, Viral; D008024:Ligands; D014759:Viral Envelope Proteins; C071440:glycoprotein H, Cytomegalovirus; C499358:glycoprotein H, Human cytomegalovirus; D012313:RNA; D012260:Ribonucleases