Exploring the effect of sequence length and composition on allele-selective inhibition of human huntingtin expression by single-stranded silencing RNAs.
Mutant huntingtin (HTT) protein is the cause of Huntington's disease (HD), an incurable neurological disorder. Almost all patients are heterozygous for mutant HTT and approaches that reduce levels of mutant HTT while leaving expression of wild-type HTT intact might be ideal options for therapeutic development. We have developed several allele-selective strategies for silencing HTT, including single-stranded silencing RNAs (ss-siRNAs). ss-siRNAs are oligonucleotides containing chemical modifications that permit action through the RNA interference (RNAi) pathway. Modified ss-siRNAs chosen to test the effects of varying oligomer length, lipid modification, the introduction of mismatched bases, and variation of chemical modification. We find that several modified ss-siRNA are potent and allele-selective inhibitors of HTT expression. An ss-siRNA with three mismatched bases relative to the CAG repeat was an allele-selective inhibitor of HTT expression in the HdhQ175 mouse model. Multiple allele-selective ss-siRNAs provide a wide platform of modifications to draw on for further optimization and therapeutic development. Our data provide insights into how ss-siRNAs can be modified to improve their properties and facilitate the discovery of the lead compounds necessary for further development.
1 Departments of Pharmacology and Biochemistry, University of Texas Southwestern Medical Center at Dallas , Dallas, Texas.
Jiaxin
J
Hu
Jing
J
Liu
Dongbo
D
Yu
Yuichiro
Y
Aiba
Suheung
S
Lee
Hannah
H
Pendergraff
Jihane
J
Boubaker
Jonathan W
JW
Artates
Clotilde
C
Lagier-Tourenne
Walt F
WF
Lima
Eric E
EE
Swayze
Thazha P
TP
Prakash
David R
DR
Corey
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D005786:Gene Expression Regulation; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D007276:Injections, Intraventricular; D008055:Lipids / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000138:chemical synthesis / Q000235:genetics* / Q000378:metabolism; D013329:Structure-Activity Relationship
C086055:HTT protein, human; D000071058:Huntingtin Protein; D008055:Lipids; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering