Cosmetic designs and products using intronic RNA

• US 9,453,219 B2
• Filed: 12/28/2007
• Issued: 09/27/2016
• Est. Priority Date: 05/15/2003
• Status: Expired due to Fees

• Current Assignee: Shi-Lung Lin
• Original Assignee: Mello Biotech Taiwan Co., Ltd.
• Inventors: Wu, David TS, Lin, Shi-Lung
• Primary Examiner(s): ANGELL, JON E
• Application Number:
  US12/003,662
• Publication Number:
  US 20090170204A1
• Time in Patent Office: 3,196 Days
• Field of Search: 435/6, 435/91.1, 435/91.31, 435/320.1, 435/455, 514/44, 536/23.1, 536/24.1, 536/24.5
• US Class Current: 1/1
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 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 Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/141 MicroRNAs, miRNAs
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 2320/50 Methods for regulating/modulating their activity

Abstract

The present invention relates to a method and composition for generating a non-naturally occurring intron and its components capable of being processed into small hairpin RNA (shRNA) and/or microRNA (miRNA) molecules by skin cells and thus inducing specific gene silencing effects on skin pigment-related genes and/or aging-causing genes in the cells. The gene silencing effects so obtained are not only useful for lightening and whitening skin colors but also useful for suppressing unwanted aging gene activities in skins.

Claims

1. An isolated nucleic acid composition for inducing microRNA mir-434-associated gene silencing effects against tyrosinase, comprising:

a SpRNAi-RGFP expression vector capable of expressing an intronic RNA molecule having a sequence which contains the sequence of SEQ.ID.NO;

8, wherein the intronic RNA molecule having the sequence of SEQ.ID.NO.8 targets tyrosinase gene.

2. The isolated nucleic acid composition as defined in claim 1, wherein said intronic RNA molecule contains a sequence which contains the sequence of SEQ ID NO:

9 for inducing gene silencing against tyrosinase and is processed into a mature microRNA having the sequence of SEQ.ID.NO.9.

3. The isolated nucleic acid composition as defined in claim 1, wherein said intronic RNA molecule contains a hairpin stem-loop structure.

4. The isolated nucleic acid composition as defined in claim 3, wherein said hairpin stem-loop structure contains a sequence which contains the sequence of either SEQ ID NO:

1 or the sequence of SEQ ID NO;

2 for inducing gene silencing against tyrosinase.

5. The isolated nucleic acid composition as defined in claim 1, which is a nucleic acid template encoding said intronic RNA molecule.

6. The isolated nucleic acid composition as defined in claim 1, further containing a 5′

-donor splice site, a 3′

-acceptor splice site, and a branch point motif.

7. The isolated nucleic acid composition as defined in claim 1, further containing sequences which contains the sequence of SEQ.ID.NO:

3, the sequence of SEQ.ID.NO;

4, and the sequence of SEQ.ID.NO;

5.

8. The isolated nucleic acid composition as defined in claim 1, further containing a recombinant mir-434-5p pre-miRNA insert having the sequence of SEQ.ID.NO:

25.

9. The isolated nucleic acid composition as defined in claim 1, which can be delivered into cells for expression.

10. The isolated nucleic acid composition as defined in claim 9, which can be delivered into cells by vector transfection.

11. The isolated nucleic acid composition as defined in claim 10, wherein said vector is a plasmid or its derivative.