Generation of species cross-reactive aptamers using "toggle" SELEX.
Species cross-reactivity facilitates the preclinical evaluation of potentially therapeutic molecules in animal models. Here we describe an in vitro selection strategy in which RNA ligands (aptamers) that bind both human and porcine thrombin were selected by "toggling" the protein target between human and porcine thrombin during alternating rounds of selection. The "toggle" selection process yielded a family of aptamers, all of which bound both human and porcine thrombin with high affinity. Toggle-25, a characteristic member, inhibited two of thrombin's most important functions: plasma clot formation and platelet activation. If appropriate targets are available, the toggle strategy is a simple measure that promotes cross-reactivity and may be generalizable to related proteins of the same species as well as to other combinatorial library screening strategies. This strategy should facilitate the isolation of ligands with needed properties for gene therapy and other therapeutic and diagnostic applications.
White
R
R
Department of Surgery, Duke University Medical Center, Durham, North Carolina 27710, USA.
Rusconi
C
C
Scardino
E
E
Wolberg
A
A
Lawson
J
J
Hoffman
M
M
Sullenger
B
B
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001777:Blood Coagulation / Q000187:drug effects; D003001:Cloning, Molecular; D020650:Combinatorial Chemistry Techniques; D003429:Cross Reactions; D015723:Gene Library; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D019151:Peptide Library; D012313:RNA / Q000138:chemical synthesis / Q000494:pharmacology*; D013045:Species Specificity; D013552:Swine; D013917:Thrombin / Q000037:antagonists & inhibitors* / Q000235:genetics; D013918:Thrombin Time
D008024:Ligands; D009838:Oligodeoxyribonucleotides; D019151:Peptide Library; D012313:RNA; D013917:Thrombin