Compositions of asymmetric interfering RNA and uses thereof
Inventors
Li, Chiang Jia • Sun, Xiangao • Rogoff, Harry • Li, Youzhi
Assignees
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Publication Number
US-11015195-B2
Publication Date
2021-05-25
Expiration Date
Abstract
The present invention provides asymmetrical duplex RNA molecules that are capable of effecting sequence-specific gene silencing. The RNA molecule comprises a first strand and a second strand. The first strand is longer than the second strand. The RNA molecule comprises a double-stranded region formed by the first strand and the second strand, and two ends independently selected from the group consisting of 5′-overhang, 3′-overhang, and blunt end. The RNA molecules of the present invention can be used as research tools and/or therapeutics.
Core Innovation
The invention relates to an asymmetric interfering RNA duplex molecule comprising an antisense strand and a sense strand. The antisense strand is longer than the sense strand and includes defined 3′-overhang and 5′-overhang lengths when duplexed with the sense strand, with sequence constraints including at least 70% complementarity to a target mRNA and a defined ribonucleotide at the last nucleotide of its 3′ end.
The sense strand consists of a defined number of nucleotides and forms a double-stranded region with the antisense strand. The double-stranded region has a defined length in base pairs, and at least the first nucleotide and the last nucleotide of the sense strand base pair with nucleotides of the antisense strand.
This structural asymmetry and constrained duplex pairing configuration define the claimed aiRNA duplex architecture. The disclosed aiRNA structure is configured to support RNA interference, including RISC incorporation, using a defined antisense complementarity profile while maintaining specific end configurations and double-stranded region constraints.
Claims Coverage
The document includes an independent claim covering an asymmetric interfering RNA (aiRNA) duplex, together with dependent claims that refine structural end configurations, duplex pairing features, and optional molecular limitations. Across the independent claim, the inventive aspects are the antisense-sense length asymmetry, independently defined 3′- and 5′-overhang lengths on the antisense strand, constrained duplex base-pair length with specific base pairing at the sense-strand termini, and antisense complementarity with a defined 3′-end ribonucleotide.
Asymmetric aiRNA duplex with longer antisense and defined overhang lengths
An asymmetric interfering RNA duplex molecule comprising an antisense strand and a sense strand, wherein the antisense strand is longer than the sense strand, includes a 3′-overhang of 1 to 9 nucleotides and a 5′-overhang of 0 to 8 nucleotides when duplexed with the sense strand, and wherein the sense strand consists of 12 to 19 nucleotides.
Defined double-stranded region length with terminal base pairing
The antisense and sense strands form a double-stranded region having a length of 11 to 19 base pairs, wherein at least the first nucleotide and last nucleotide of the sense strand base pair with nucleotides of the antisense strand.
Antisense complementarity to target mRNA with defined 3′-end ribonucleotide
The antisense strand is at least 70% complementary to a target mRNA and has the last nucleotide of its 3′ end consisting of an A, U, G or C ribonucleotide.
Refined duplex base-pairing fidelity for the double-stranded region
The RNA duplex molecule is characterized by a double-stranded region made of perfectly complementary sequences.
Refined overhang length combinations for the antisense strand
The RNA duplex molecule includes antisense strand 5′- and 3′-overhangs characterized by specified overhang lengths of 2, 3, or 4 nucleotides.
GC content constraint for the double-stranded region
The RNA duplex molecule includes a double-stranded region whose GC content is 30% to 50%.
Modified nucleotide or analogue inclusion
The claimed RNA duplex molecule further includes at least one modified nucleotide or an analogue.
Therapeutic use for treating or preventing cancer or a tumor by administration
A method for treating or preventing cancer or a tumor by administering an RNA duplex molecule as defined in claim 1 to a subject in need.
Overall, the claim set centers on a specific asymmetric aiRNA duplex architecture: a longer antisense strand with independently defined 3′- and 5′-overhang lengths, a constrained-length double-stranded region with terminal base pairing, and an antisense strand that is at least 70% complementary to a target mRNA with a specified 3′-end ribonucleotide. Dependent claims further narrow duplex composition and base-pairing characteristics, refine permissible antisense overhang length combinations, optionally add modified nucleotides or analogues, and extend the duplex to methods of treating or preventing cancer or tumor via administration.
Stated Advantages
The disclosed aiRNA structure is configured to support RNA interference, including RISC incorporation.
The invention addresses the need for sequence-specific gene silencing in eukaryotic cells while controlling undesirable biological effects associated with RNA interference.
The specific end configurations and double-stranded region constraints are intended to improve performance.
Documented Applications
RNA interference in eukaryotic cells.
Treating or preventing cancer or a tumor by administration of the RNA duplex molecule.
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