RNA-splicing and processing-directed gene silencing and the relative applications thereof
OwnerShi-Lung Lin, Shao-Yao Yinginventors
Life of this patent
Estimating the term…
Abstract
The present invention relates to a method for generating a recombinant gene composition, which is able to elicit specific gene silencing effects through RNA splicing and/or processing mechanisms, and the relative utilization thereof. The recombinant gene molecule so generated is useful not only for delivering desirable gene function into the transfected cells thereof but also for suppressing undesirable gene function in the transfected cells, respectively or simultaneously. Furthermore, the derivative products of this novel recombinant gene have multiple utilities in probing gene function, validating drug target, and treating as well as preventing gene-related diseases.
Claims57
A method for inducing of RNA splicing/processing-associated gene silencing effects comprises the steps of:
- (a) Constructing a recombinant nucleic acid composition containing at least an intron flanked with a plurality of exons, wherein said intron can be cleaved out of the exons by RNA splicing and/or processing for gene silencing and said exons can be linked together to form a gene with desired function;
- (b) Cloning said recombinant nucleic acid composition into an expression-competent vector;
- (c) Introducing said vector into a cell, cells, tissue or in vivo;
- (d) Generating RNA transcript of said recombinant nucleic acid composition;
- and
- (e) Releasing the metabolic products of said intron by RNA splicing/processing mechanisms, so as to provide gene silencing effects against the genes containing sequences homologous to said intron.
A method for inducing of posttranscriptional gene silencing effects comprises:
- (a) Constructing a recombinant gene composition containing a functional RNA polymerase promoter and at least an intron flanked with a plurality of exons, wherein said intron can be cleaved out of the exons by RNA splicing and/or processing for gene silencing and said exons can be linked to form a gene with desired function;
- (b) Introducing said recombinant gene composition into a cell or in vivo;
- (c) Generating RNA transcript of said recombinant gene composition;
- and
- (d) Releasing the metabolic products of said intron by RNA splicing/processing mechanisms, so as to provide gene silencing effects against the genes containing sequences homologous to said intron.
A recombinant nucleic acid composition for inducing of RNA splicing/processing-associated gene silencing comprises:
- (a) At least an intron, wherein said intron is flanked with a plurality of exons and can be cleaved out of the exons by cellular RNA splicing and/or processing machinery;
- and
- (b) A plurality of exons, wherein said exons can be linked to form a gene possessing desired function.
An insert-containing intron composition, wherein the insert of said intron composition can be inserted into the intron area of a gene for producing of a desired RNA molecule through RNA splicing/processing mechanisms, comprises:
- (a) An insert;
- (b) A splicing donor site;
- (c) A splicing acceptor site;
- (d) A branch point domain for splicing recognition;
- (e) At least a poly-pyrimidine tract for spliceosome interaction;
- and
- (f) A plurality of nucleic acid linkers for connection of the above components.
39 The composition as defined in claims 37 and 38, wherein said insert is a nucleic acid sequence containing components and/or analogs either homologous or complementary to at least a targeted gene selected from the group consisting of pathogenic nucleic acids, viral genes, mutated genes, oncogenes and many other types of functional as well as non-functional genes.
Dependencies are inferred from the claim text.