Rational design of antisense oligonucleotides targeting single nucleotide polymorphisms for potent and allele selective suppression of mutant Huntingtin in the CNS
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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Rational design of antisense oligonucleotides targeting single nucleotide polymorphisms for potent and allele selective suppression of mutant Huntingtin in the CNS. / Østergaard, Michael E; Southwell, Amber L; Kordasiewicz, Holly; Watt, Andrew T; Skotte, Niels H; Doty, Crystal N; Vaid, Kuljeet; Villanueva, Erika B; Swayze, Eric E; Bennett, C Frank; Hayden, Michael R; Seth, Punit P.
I: Nucleic Acids Research, Bind 41, Nr. 21, 11.2013, s. 9634-50.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Rational design of antisense oligonucleotides targeting single nucleotide polymorphisms for potent and allele selective suppression of mutant Huntingtin in the CNS
AU - Østergaard, Michael E
AU - Southwell, Amber L
AU - Kordasiewicz, Holly
AU - Watt, Andrew T
AU - Skotte, Niels H
AU - Doty, Crystal N
AU - Vaid, Kuljeet
AU - Villanueva, Erika B
AU - Swayze, Eric E
AU - Bennett, C Frank
AU - Hayden, Michael R
AU - Seth, Punit P
PY - 2013/11
Y1 - 2013/11
N2 - Autosomal dominant diseases such as Huntington's disease (HD) are caused by a gain of function mutant protein and/or RNA. An ideal treatment for these diseases is to selectively suppress expression of the mutant allele while preserving expression of the wild-type variant. RNase H active antisense oligonucleotides (ASOs) or small interfering RNAs can achieve allele selective suppression of gene expression by targeting single nucleotide polymorphisms (SNPs) associated with the repeat expansion. ASOs have been previously shown to discriminate single nucleotide changes in targeted RNAs with ∼5-fold selectivity. Based on RNase H enzymology, we enhanced single nucleotide discrimination by positional incorporation of chemical modifications within the oligonucleotide to limit RNase H cleavage of the non-targeted transcript. The resulting oligonucleotides demonstrate >100-fold discrimination for a single nucleotide change at an SNP site in the disease causing huntingtin mRNA, in patient cells and in a completely humanized mouse model of HD. The modified ASOs were also well tolerated after injection into the central nervous system of wild-type animals, suggesting that their tolerability profile is suitable for advancement as potential allele-selective HD therapeutics. Our findings lay the foundation for efficient allele-selective downregulation of gene expression using ASOs-an outcome with broad application to HD and other dominant genetic disorders.
AB - Autosomal dominant diseases such as Huntington's disease (HD) are caused by a gain of function mutant protein and/or RNA. An ideal treatment for these diseases is to selectively suppress expression of the mutant allele while preserving expression of the wild-type variant. RNase H active antisense oligonucleotides (ASOs) or small interfering RNAs can achieve allele selective suppression of gene expression by targeting single nucleotide polymorphisms (SNPs) associated with the repeat expansion. ASOs have been previously shown to discriminate single nucleotide changes in targeted RNAs with ∼5-fold selectivity. Based on RNase H enzymology, we enhanced single nucleotide discrimination by positional incorporation of chemical modifications within the oligonucleotide to limit RNase H cleavage of the non-targeted transcript. The resulting oligonucleotides demonstrate >100-fold discrimination for a single nucleotide change at an SNP site in the disease causing huntingtin mRNA, in patient cells and in a completely humanized mouse model of HD. The modified ASOs were also well tolerated after injection into the central nervous system of wild-type animals, suggesting that their tolerability profile is suitable for advancement as potential allele-selective HD therapeutics. Our findings lay the foundation for efficient allele-selective downregulation of gene expression using ASOs-an outcome with broad application to HD and other dominant genetic disorders.
KW - Alleles
KW - Animals
KW - Base Pairing
KW - Brain
KW - Cells, Cultured
KW - Down-Regulation
KW - Fluorine
KW - Humans
KW - Huntington Disease
KW - Mice
KW - Mice, Transgenic
KW - Mutation
KW - Nerve Tissue Proteins
KW - Oligonucleotides, Antisense
KW - Polymorphism, Single Nucleotide
KW - Rats
KW - Rats, Sprague-Dawley
KW - Ribonuclease H
U2 - 10.1093/nar/gkt725
DO - 10.1093/nar/gkt725
M3 - Journal article
C2 - 23963702
VL - 41
SP - 9634
EP - 9650
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 21
ER -
ID: 153451258