Potent and nontoxic antisense oligonucleotides containing locked nucleic acids.
Insufficient efficacy and/or specificity of antisense oligonucleotides limit their in vivo usefulness. We demonstrate here that a high-affinity DNA analog, locked nucleic acid (LNA), confers several desired properties to antisense agents. Unlike DNA, LNA/DNA copolymers were not degraded readily in blood serum and cell extracts. However, like DNA, the LNA/DNA copolymers were capable of activating RNase H, an important antisense mechanism of action. In contrast to phosphorothioate-containing oligonucleotides, isosequential LNA analogs did not cause detectable toxic reactions in rat brain. LNA/DNA copolymers exhibited potent antisense activity on assay systems as disparate as a G-protein-coupled receptor in living rat brain and an Escherichia coli reporter gene. LNA-containing oligonucleotides will likely be useful for many antisense applications.
Wahlestedt
C
C
Center for Genomics Research and Department of Neuroscience, Karolinska Institutet, S-171 77 Stockholm, Sweden. claes.wahlestedt@cgr.ki.se
Salmi
P
P
Good
L
L
Kela
J
J
Johnsson
T
T
Hökfelt
T
T
Broberger
C
C
Porreca
F
F
Lai
J
J
Ren
K
K
Ossipov
M
M
Koshkin
A
A
Jakobsen
N
N
Skouv
J
J
Oerum
H
H
Jacobsen
M H
MH
Wengel
J
J
D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D001943:Breast Neoplasms; D002421:Caudate Nucleus / Q000187:drug effects; D002552:Cerebral Ventricles / Q000187:drug effects / Q000502:physiology; D015195:Drug Design; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D020319:Oligodeoxyribonucleotides, Antisense / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology*; D011699:Putamen / Q000187:drug effects; D051381:Rats; D017465:Receptors, Opioid, delta / Q000235:genetics; D016914:Ribonuclease H / Q000187:drug effects / Q000378:metabolism; D013238:Stereotaxic Techniques; D013873:Thionucleotides; D014407:Tumor Cells, Cultured
D020319:Oligodeoxyribonucleotides, Antisense; D017465:Receptors, Opioid, delta; D013873:Thionucleotides; D016914:Ribonuclease H