Unigene unidirectional antisense library
AssigneeWelgene Pharmaceuticals, Inc.
InventorsJong-Gu Park, Ik-Jae Moon, Yun-Han Lee
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Abstract
The present invention provides a high throughput system for functional genomics using a unigene antisense library comprising LC-antisense compounds. The antisense compounds were specific and effective for the elimination of target mRNA. Thus, the system of the present invention is used as temporary knock-down system to unveil functions of genes critical for diseases. The system of the present invention can be adopted not only for functional genomics but also for effectively validating target for antisense or other molecular therapeutics against various malignancies, infections, and other diseases by blocking specific genes involved in the disease.
Claims31
A library of a multitude of unique single-stranded nucleic acids, said library comprising:
- a multiplicity of compartments, each of said compartments comprising one or more single-stranded LC-antisense compound derived from recombinant bacteriophage or phagemid vector comprising at least one unique unidirectional antisense nucleic acid insert in an aqueous medium,
- wherein said LC-antisense compound is capable of being introduced into a host cell, and which is capable of specifically binding to a nucleic acid in said host cell that is substantially complementary to said unique antisense nucleic acid insert.
A method of making a library comprising a multitude of unique single-stranded nucleic acids, which comprises one or more single-stranded LC-antisense compound derived from recombinant bacteriophage or phagemid vector comprising at least one unique unidirectional antisense nucleic acid insert, comprising:
- inserting a nucleic acid fragment unidirectionally into said bacteriophage or phagemid vector by unidirectionally cloning the nucleic acid fragments into said vector.
A method for specifically inhibiting growth of liver cancer cells, comprising administering to said cells a large circular antisense compound targeted to polymyositis/scleroderma autoantigen, ESTs (N21972), Nuclear matrix protein p84, Gamma-aminobutyric acid (GABA) A receptor beta 3, SRY (sex-determining region Y)-box 9, ESTs (H13112), ESTs (AW294133), Primase, polypeptide 1 (49 kD), Human EV12 protein gene, epidermal growth factor receptor pathway substrate 8 or protein tyrosine phosphatase, non-receptor type 2.
A method for specifically inhibiting growth of lung cancer cells, comprising administering to said cells a large circular antisense compound targeted to TGF-β stimulated protein, TSC-22, General transcription factor II H, Cytochrome P450, subfamily III A, polypeptide 7, KIAA0094 protein (D42084), MAX dimerization protein, Serine/treonine kinase 13 (aurora/IPL 1-kike), ESTs (AI057094), Ras-related GTP-binding protein, MHC class I region ORF, or Tumor necrosis factor receptor superfamily, member 7.
A high throughput system for conducting a functional genomics assay with a unigene unidirectional antisense library comprising the steps of:
- (i) forming large circular antisense molecule-carrier complexes with said unigene unidirectional antisense library;
- (ii) transfecting the complexes into host cells to eliminate endogenously expressed substantially complementary transcripts;
- (iii) screening for a change in phenotype of the host cell;
- (iv) identifying the gene that caused the change in phenotype in (iii).
A high throughput system for conducting massive functional genomics assays, which is performed by applying a unigene unidirectional antisense library to a cell line of a particular disease comprising the following steps:
- 1) making an antisense library by massively parallel production of LC-antisense compounds to a large number unigenes;
- 2) plating a population of host cells in multi-well plates;
- 3) forming an LC-antisense compound-carrier complex with the antisense library of step 1);
- 4) performing primary gene functional analysis by transfection of the complex of step 3) into the population of host cells;
- and
- 5) performing additional functional analysis of the gene screened in step 4).
A high throughput system for massive functional genomics performed by applying a macroarray or microarray assembly to disease cells comprising the steps of:
- 1) making an antisense array by selecting unigene LC-antisense compounds;
- 2) plating a population of host cells in multi-well plates;
- 3) forming LC-antisense compound-carrier complexes on the antisense array of step 1);
- 4) performing primary gene functional analysis by transfection of the complexes of step 3) into the population of cells;
- and
- 5) performing additional functional assays of the genes screened in step 4).
Dependencies are inferred from the claim text.