Methods and compositions for the production of siRNAs

Patent metadata

Field	Value
Publication number	US-9840703-B2
Application number	US-201414758924-A
Country	US
Kind code	B2
Filing date	Jan 9, 2014
Priority date	Jan 11, 2013
Publication date	Dec 12, 2017
Grant date	Dec 12, 2017

Abstract

The technology described herein relates to siRNAs, e.g., methods and compositions relating to the production of siRNAs in bacterial cells.

First claim

What is claimed herein is:

1. A bacterial cell comprising a viral siRNA-binding polypeptide and a dsRNA comprising an exogenous nucleic acid sequence substantially complementary to a target RNA.

2. The bacterial cell of claim 1 , wherein the siRNA-binding polypeptide comprises a purification tag.

3. The bacterial cell of claim 1 , wherein the siRNA-binding polypeptide is encoded by a nucleic acid.

4. The bacterial cell of claim 1 , wherein the siRNA-binding polypeptide is selected from the group consisting of:
p19 polypeptide; tombusvirus p19 polypeptide; B2 polypeptide; HC-Pro polypeptide; p38 polypeptide; p122 polypeptide; p130 polypeptide; p21 polypeptide; p1b polypeptide; and NS3 polypeptide.

5. The bacterial cell of claim 1 , wherein the dsRNA is greater than 21 nucleotides in length.

6. The bacterial cell of claim 1 , wherein the dsRNA is a hairpin RNA.

7. The bacterial cell of claim 1 , wherein the bacterial cell expresses an RNase III polypeptide.

8. The bacterial cell of claim 1 , wherein the bacterial cell expresses an RNase III polypeptide encoded by an exogenous nucleic acid sequence.

9. The bacterial cell of claim 1 , wherein the bacterial cell is an Escherichia coli cell.

10. The bacterial cell of claim 1 , wherein at least one of the siRNA-binding polypeptide and the dsRNA are constitutively expressed.

11. The bacterial cell of claim 1 , wherein at least one of the siRNA-binding polypeptide and the dsRNA are inducibly expressed.

12. The bacterial cell of claim 1 , wherein the DNA encoding at least one of the siRNA-binding polypeptide or the dsRNA is part of a plasmid.

13. The bacterial cell of claim 1 , wherein the dsRNA comprises nucleic acid sequences substantially complementary to a multiplicity of target RNAs.

Assignees

• Children'S Medical Center Corp

Inventors

• Lieberman Judy
• Huang Linfeng

Classifications

• A61K31/713

  Double-stranded nucleic acids or oligonucleotides · CPC title

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N2330/50

  Biochemical production, i.e. in a transformed host cell · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/111 Primary

  General methods applicable to biologically active non-coding nucleic acids · CPC title