Composition for producing microRNA precursors as drugs for enhancing wound healing and production method of the microRNA precursors

• US 10,196,662 B2
• Filed: 01/20/2015
• Issued: 02/05/2019
• Est. Priority Date: 08/10/2012
• Status: Active Grant

• Current Assignee: Mello Biotechnology, Inc.
• Original Assignee: Mello Biotechnology, Inc.
• Inventors: Wu, David T S, Lin, Shi-Lung, Chen, Jack S. K.
• Primary Examiner(s): Nguyen, Quang
• Application Number:
  US14/601,087
• Publication Number:
  US 20150132805A1
• Time in Patent Office: 1,477 Days
• CPC Class Codes: C12N 1/38 Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound C12N1/34 takes precedence
• CPC Class Codes: C12N 15/635 Externally inducible repressor mediated regulation of gene expression, e.g. tetR inducible by tetracyline
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides

Abstract

This invention generally relates to a composition and its production method useful for developing drugs and/or therapies for enhancing wound healing, in particular scarless wound healing. Particularly, the present invention teaches the essential processes necessary for producing and purifying embryonic stem cell (ESC)-specific RNA compositions, such as messenger RNAs (mRNA), microRNA precursors (pre-miRNA), and small hairpin RNAs (shRNA), which are useful for treating human diseases and lesions.

Claims

1. A method for enhancing wound healing in vivo using hairpin-like microRNA precursor (pre-miRNA), comprising:

(a) producing the hairpin-like microRNA precursor in Escherichia coli (E. coli) strain competent cells having at least an expression vector encoding the hairpin-like microRNA precursor comprising SEQ ID NO;

3 operably linked to a CMV promoter or a eukaryotic EF1alpha promoter that is induced by a chemical agent containing 3-morpholinopropane-1-sulfonic acid (MOPS);

(b) providing at least a hairpin-like microRNA precursor comprising SEQ ID NO;

3 that is produced in (a);

(c) providing at least a wounded or damaged tissue cell in vivo; and

(d) contacting the hairpin-like microRNA precursor of (b) with the at least a wounded or damaged tissue cell of (c), so as to induce a wound healing effect of the hairpin-like microRNA precursor on the at least one wounded or damaged tissue cell.

2. The method as defined in claim 1, wherein said expression vector is pLenti-EF1alpha-RGFP-miR302.

3. The method as defined in claim 1, wherein said hairpin-like microRNA precursor comprises a hairpin-like sequence of SEQ ID NO:

8.

4. The method as defined in claim 1, wherein said hairpin-like microRNA precursor is a miR-302 precursor (pre-miR-302).

5. The method as defined in claim 4, wherein said pre-miR-302 comprises a sequence of miR-302a.

6. The method as defined in claim 4, wherein said pre-miR-302 is capable of being used as a part of drug ingredients for pharmaceutical and therapeutic applications.

7. The method as defined in claim 1, wherein said hairpin-like microRNA precursor is capable of being used as a part of drug ingredients for pharmaceutical and therapeutic applications.

8. The method as defined in claim 1, wherein the hairpin-like microRNA precursor is capable of treating wounds in mammals.