Una oligomers having reduced off-target effects in gene silencing

a technology of una oligomers and gene silencing, applied in the field of gene silencing techniques, can solve the problems of difficult to carry out, rna interference is the occurrence of off-target effects, and the silencing technique is difficult to achieve, so as to reduce ttr and off-target effects

Inactive Publication Date: 2015-10-29
ARCTURUS THERAPEUTICS
View PDF4 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0007]In particular, the active agents are UNA oligomers having reduced off-target effects in gene silencing.
[0009]The UNA oligomers of this invention can be effective for gene silencing with reduced off-target effects for a wide range of gene targets.
[0011]In some aspects, this invention provides therapeutics for amyloidosis. More particularly, this invention relates to methods for treating transthyretin-related amyloidosis with UNA oligomers having reduced off-target effects in knockdown of transthyretin.
[0015]A UNA oligomer for inhibiting expression of a target gene, the oligomer comprising a first strand and a second strand, each of the strands being 19-29 monomers in length, the monomers comprising UNA monomers and nucleic acid monomers, wherein the oligomer has a duplex structure of from 14 to 29 monomers in length, and wherein the oligomer has reduced off-target effects as compared to a siRNA with the same target.
[0019]The UNA oligomer above, wherein the UNA oligomer has one or two overhangs. The UNA oligomer above, wherein the UNA oligomer inhibits TTR expression with reduced off-target effects. The UNA oligomer above, wherein the UNA oligomer inhibits apolipoprotein gene expression with reduced off-target effects.
[0033]The method above, wherein the TTR-related amyloidosis is ATTR. The method above, wherein the subject is human. The method above, wherein the method reduces TTR in the subject with off-target effects reduced by at least 10% as compared to control.

Problems solved by technology

A major drawback of gene silencing techniques such as RNA interference is the occurrence of off-target effects.
Off-target effects occur when a gene-silencing agent has an effect on a gene product to which it is not targeted, and is therefore unwanted.
Off-target effects loom as one of the main hurdles for developing gene silencing therapeutics.
These conventional methods have been able to reduce off-target effects, however, they can be difficult to carry out.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Una oligomers having reduced off-target effects in gene silencing
  • Una oligomers having reduced off-target effects in gene silencing
  • Una oligomers having reduced off-target effects in gene silencing

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0169]This example shows that UNA oligomers dramatically reduce off target activity of the passenger strand in gene silencing by RNA interference. The reduction in passenger strand off target activity can depend on the positioning of various UNA monomers in the oligomer. In this example, it is shown that the presence of a combination of UNA monomers in three positions in a UNA oligomer, more specifically, in the passenger strand at the 5′ end and at the 3′ end, as well as in the guide strand at the 3′ end, greatly reduced off target knockdown activity by the passenger strand.

[0170]UNA oligomers targeted to ApoCIII having reduced off-target effects are shown in Table 3. As used herein, a duplex oligomer is represented with the passenger strand above, and the guide strand below. The end group numbering will depend on the identity of the terminal monomer, as described above.

TABLE 3UNA oligomers ATX98 and ATX100SEQ IDNO:OLIGOMER33ATX1-ÃAAAGGGACAGUAUUCUCAÛmU-3′34983′-mUÛUUUUCCCUGUCAUAAGA...

example 2

[0175]FIG. 3 shows that certain UNA oligomers had at least comparable knockdown levels of activity to conventional siRNAs for TTR mRNA expression. ATX13, ATX14, ATX15, ATX16, ATX17, ATX21 and ATX25 were targeted to the 3′-UTR of human TTR, and therefore were targeted to both wild-type V30V and V30M mutant TTR.

[0176]In particular, UNA oligomer ATX13 having a UNA monomer in the first strand located at the 1 (5′) end, and UNA oligomer ATX15 having a UNA monomer in the first strand located at the 1 (5′) end and two UNA monomers in the second strand located at the 3 (3′) end in the 20th and 21st positions counting from the 5′ end, had at least comparable knockdown levels of activity as compared to conventional siRNA ATS-91.

[0177]Further, UNA oligomer ATX21 having a UNA monomer in the first strand located at the 1 (5′) end, one UNA monomer in the first strand located at the 3 (3′) end in the 20th position counting from the 5′ end, and one UNA monomer in the second strand located at the 3 ...

example 3

[0180]FIG. 5 shows the protocol for measuring off-target (OT) effects. A Luciferase reporter assay using PSICHECK vector was established. For each measurement, 4 plasmids were constructed: guide strand GSCM and guide strand GSSM for second strand or antisense knockdown, and passenger strand PSCM and passenger strand PSSM for first strand or sense strand knockdown. The reporter plasmid was co-transfected with UNA oligomer into HeLa cells. In this system, if the UNA oligomer binds to the target sequence inserted in 3-UTR of luciferase, then the chemiluminescent signal is reduced or disappeared.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
lengthaaaaaaaaaa
sizeaaaaaaaaaa
concentrationaaaaaaaaaa
Login to view more

Abstract

This invention provides UNA oligomers for gene silencing with reduced off-target effects. The UNA oligomers can have a first strand and a second strand, each of the strands being 19-29 monomers in length, the monomers being UNA monomers and various nucleic acid monomers. Embodiments include pharmaceutical compositions and methods for treating or preventing TTR-related amyloidosis with reduced off-target effects by administering a UNA oligomer to a subject.

Description

SEQUENCE LISTING[0001]This application includes a Sequence Listing submitted electronically herewith as an ASCII file created on Mar. 24, 2015, named ARC1246WO_SL.txt, which is 30,972 bytes in size, and is hereby incorporated by reference in its entirety.BACKGROUND OF THE INVENTION[0002]A major drawback of gene silencing techniques such as RNA interference is the occurrence of off-target effects. Off-target effects occur when a gene-silencing agent has an effect on a gene product to which it is not targeted, and is therefore unwanted. Off-target effects loom as one of the main hurdles for developing gene silencing therapeutics.[0003]One method to reduce off-target effects is to intelligently design the structure of the gene silencing agent to avoid effects on genes other than the desired target. In some cases, the gene silencing agents could be pooled so that the concentration of agents having a particular off-target effect would be reduced. In other cases, the gene silencing agent ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/113A61K47/14A61K31/713
CPCC12N15/113A61K31/713C12N2310/32C12N2310/14A61K47/14C12N2310/323A61P25/00A61P25/02A61P25/28A61P43/00A61P5/14A61P9/00
Inventor TACHIKAWA, KIYOSHIPAYNE, JOSEPH E.CHIVUKULA, PADMANABH
Owner ARCTURUS THERAPEUTICS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap