Abstract: A computer system (such as a controller) that selects channels and/or channel widths for use during communication in a shared band of frequencies is described. During operation, the computer system may receive, associated with access points in a region (such as a zone), information specifying unavailable channels associated with the shared band of frequencies, where the unavailable channels are currently used by the access points. For example, the information may be included in access-point status reports from the access points. Then, based at least in part on the unavailable channels, the computer system may determine the channels and/or the channel widths for use by the access points during communication in the shared band of frequencies in the region. Next, the computer system may provide, addressed to the access points, second information specifying the determined channels and/or the channel width.

Abstract: This invention generally relates to a method for developing, generating and selecting human embryonic stem (hES)-like pluripotent cells using transgenic expression of intronic microRNA-like RNA agents. More particularly, the present invention relates to a method and composition for generating a non-naturally occurring intron and its intronic components capable of being processed into mir-302-like RNA molecules in mammalian cells and thus inducing certain specific gene silencing effects on differentiation-related and fate-determinant genes of the cells, resulting in reprogramming the cells into a pluripotent embryonic stem (ES)-cell-like state. The ES-like cells so obtained are strongly express hES cell markers, such as Oct3/4, SSEA-3 and SSEA-4, and can be guided into various tissue cell types by treating certain hormones and/or growth factors under a feeder-free cell culture condition in vitro, which may be used for transplantation and gene therapies.

Abstract: An isolated RNA comprising an intron RNA that is released in a cell, thereby modulating the function of a target gene. Also disclosed are a composition comprising a chemokine and an isolated RNA of the invention or a DNA template for the isolated RNA, a composition comprising one or more agents that induce RNA-mediated modulation of the functions of two or more target genes in a cell, and methods of using these compositions for modulating the functions of genes in a cell.

Abstract: Patterns of microRNA (miRNA) expression are correlated to the degrees of tumor cell differentiation in human prostate cancer. MiRNAs can complementarily bind to either oncogenes or tumor suppressor genes, resulting in targeted gene silencing and thus changes of cellular tumorigenecity. Using miRNA microarray analysis, 8 down-regulated and 3 up-regulated known miRNAs in androgen-independent human prostate cancer cell lines, such as LNCaP C4-2B and PC3, compared to those androgen-dependent cell lines, such as LNCaP and PC3-AR9 were consistently detected. Fluorescent in-situ hybridization assays in human prostate cancer tissue arrays containing sixty patients at different stages also showed the same miRNA expression patterns in hormone-refractory prostate carcinomas (HRPC) compared to androgen-sensitive non-cancerous prostate epithelium. In-vitro tumorigenecity assays using one of the identified miRNAs, mir-146a, were performed to provide validation of its function in prostate cancer.

Abstract: An isolated RNA comprising an intron RNA that is released in a cell, thereby modulating the function of a target gene. Also disclosed are a composition comprising a chemokine and an isolated RNA of the invention or a DNA template for the isolated RNA, a composition comprising one or more agents that induce RNA-mediated modulation of the functions of two or more target genes in a cell, and methods of using these compositions for modulating the functions of genes in a cell.

Abstract: The invention relates to isolated nucleic acids comprising mir-302 genes. Also disclosed are expression vectors, host cells, and transgenic animals containing the nucleic acids, and use of the nucleic acids to generate ES-like cells.

Abstract: The present invention provides novel compositions and methods for suppressing the function or activity of a targeted gene through a novel intracellular piRNA-mediated RNAi mechanism, using RNA-DNA duplex constructs. The invention further provides novel methods and compositions for generating or producing RNA-DNA duplex agents, whose quantity is high enough to be used for the invention's gene silencing transfection and possibly in therapeutics applications. This improved RNA-polymerase chain reaction (RNA-PCR) method utilizes thermocycling steps of promoter-linked DNA or RNA template synthesis, in vitro transcription and then reverse transcription to bring up the amount of RNA-DNA duplexes up to two thousand folds within one round of the above procedure for using in D-RNAi-directed gene silencing.

Abstract: The present invention provides novel compositions and methods for suppressing the function or activity of a targeted gene through a novel intracellular piRNA-mediated RNAi mechanism, using RNA-DNA duplex constructs. The invention further provides novel methods and compositions for generating or producing RNA-DNA duplex agents, whose quantity is high enough to be used for the invention's gene silencing transfection and possibly in therapeutics applications. This improved RNA-polymerase chain reaction (RNA-PCR) method utilizes thermocycling steps of promoter-linked DNA or RNA template synthesis, in vitro transcription and then reverse transcription to bring up the amount of RNA-DNA duplexes up to two thousand folds within one round of the above procedure for using in D-RNAi-directed gene silencing.

Abstract: Patterns of microRNA (miRNA) expression are correlated to the degrees of tumor cell differentiation in human prostate cancer. MiRNAs can complementarily bind to either oncogenes or tumor suppressor genes, resulting in targeted gene silencing and thus changes of cellular tumorigenecity. Using miRNA microarray analysis, 8 down-regulated and 3 up-regulated known miRNAs in androgen-independent human prostate cancer cell lines, such as LNCaP C4-2B and PC3, compared to those androgen-dependent cell lines, such as LNCaP and PC3-AR9 were consistently detected. Fluorescent in-situ hybridization assays in human prostate cancer tissue arrays containing sixty patients at different stages also showed the same miRNA expression patterns in hormone-refractory prostate carcinomas (HRPC) compared to androgen-sensitive non-cancerous prostate epithelium. In-vitro tumorigenecity assays using one of the identified miRNAs, mir-146a, were performed to provide validation of its function in prostate cancer.

Abstract: This invention generally relates to a method for developing, generating and selecting human embryonic stem (hES)-like pluripotent cells using transgenic expression of intronic microRNA-like RNA agents. More particularly, the present invention relates to a method and composition for generating a non-naturally occurring intron and its intronic components capable of being processed into mir-302-like RNA molecules in mammalian cells and thus inducing certain specific gene silencing effects on differentiation-related and fate-determinant genes of the cells, resulting in reprogramming the cells into a pluripotent embryonic stem (ES)-cell-like state. The ES-like cells so obtained are strongly express hES cell markers, such as Oct3/4, SSEA-3 and SSEA-4, and can be guided into various tissue cell types by treating certain hormones and/or growth factors under a feeder-free cell culture condition in vitro, which may be used for transplantation and gene therapies.

Abstract: The present invention provides novel compositions and methods for suppressing the function or activity of a targeted gene through a novel intracellular piRNA-mediated RNAi mechanism, using RNA-DNA duplex constructs. The invention further provides novel methods and compositions for generating or producing RNA-DNA duplex agents, whose quantity is high enough to be used for the invention's gene silencing transfection and possibly in therapeutics applications. This improved RNA-polymerase chain reaction (RNA-PCR) method utilizes thermocycling steps of promoter-linked DNA or RNA template synthesis, in vitro transcription and then reverse transcription to bring up the amount of RNA-DNA duplexes up to two thousand folds within one round of the above procedure for using in D-RNAi-directed gene silencing.

Abstract: The present invention relates to a method and composition for generating an artificial intron and its components capable of producing microRNA (miRNA) molecules and thus inducing specific gene silencing effects through intracellular RNA interference (RNAi) mechanisms, and the relative utilization thereof. The miRNA-producing intron so generated is not only useful for delivering desired miRNA function into the intron-mediated transgenic organisms or cells but also useful for suppressing unwanted gene function in the transgenic organisms or cells thereof. Furthermore, the derivative products of this novel man-made miRNA-producing intron have utilities in probing gene functions, validating drug targets, generating transgenic animals and gene-modified plants, developing anti-viral vaccines and treating as well preventing gene-related diseases (gene therapy).

Abstract: The present invention provides methods of immobilizing proteins onto a support, using a cellular expression system or a cell-free expression system, to attach a ligand to the C-terminus of the protein, by an intein-mediated or a puromycin-mediated approach. A method for improving the efficiency of intein-mediated ligand attachment is also provided. The methods of the present invention are useful in the preparation of protein microarrays.

Abstract: An in vivo method for labelling a protein having an N-terminal cysteine is disclosed. A probe comprising a thioester group and a detectable label is introduced into a cell expressing the N-terminal cysteine protein, allowing for the N-terminal cysteine to cleave the thioester bond and resulting in the label being covalently attached to the N-terminus of the protein by an amide bond.

Abstract: An isolated RNA comprising an intron RNA that is released in a cell, thereby modulating the function of a target gene. Also disclosed are a composition comprising a chemokine and an isolated RNA of the invention or a DNA template for the isolated RNA, a composition comprising one or more agents that induce RNA-mediated modulation of the functions of two or more target genes in a cell, and methods of using these compositions for modulating the functions of genes in a cell.

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.

Abstract: The present invention provides a fast, simple and reliable polymerase thermocycling reaction procedure for the linear amplification of nucleic acid sequences from cellular RNAs or genomes or both. The principle of this polymerase thermocycling reaction method relies upon the thermal cycling steps of promoter-linked nucleic acid template synthesis and in-vitro transcriptional amplification to bring up the amount of desired nucleic acid sequences up to two thousand fold within one cycle of the above procedure. Neither RNase H activity nor alkaline degradation is used in order to protect the RNA sequences of resulting amplified products which are ready for further applications, such as genechip/microarray analysis, in-vitro translation reaction and induction of RNA interference. Without the preferential amplification drawback of PCR/RT-PCR methods, the accuracy and resolution of current molecular biology analysis and diagnosis can be significantly improved by the present invention.

Abstract: The present invention provides a fast, simple and specific method for generating a complete full-length cDNA library from single cells. The first reverse transcription of intracellular mRNAs with an oligo(dT)n-promoter primer introduces a recognition site for following transcription of newly reverse-transcribed cDNAs. The poly-nucleotide tailing of above cDNAs in addition to aforementioned promoter region further forms binding templates for specific PCR amplification. After repeating the reverse transcription, transcription, reverse transcription and PCR procedure, we can multiply a single copy of mRNA to two billion folds by calculation based upon the comparison between the amount of a synthesized cDNA library and that of theoretically presumed mRNAs within a cell (0.1 pg). In conjunction with a cell fixation and permeabilization step, the complete full-length cDNA library can be directly generated from few single cells without mRNA degradation.

Abstract: The present invention provides a simple, specific and nontoxic gene knock-out method by formation of covalently bonding between modified probes and targeted sequences. When covalently modified first strand probes are hybridized with a second strand of targeted gene transcripts, certain modified bases of said first strand will interact with natural bases of said second strand to form covalent bonds by which the translation of said second strand is inhibited. Because the hybridization of said two strands generates covalent base-pairing only between their complementary homology region(s), such specificity increases the targeting efficiency of a gene knock-out system.

Abstract: Excess amount of modified subtracter DNA from control cells is generated by carboxylating the base structures of its certain nucleotides with chemical agents in order to introduce covalent affinity between the modified subtracter and a non-modified tester DNA. Hybridization of the control subtracter and the experimental tester DNA is performed with a heat-melting and then cool-reassociation technique. While the desired different (heterologous) sequences remain in the form of hydrogen-binding, common (homologous) sequences of the hybridized DNA are covalently bonded to each other. Since the covalent bonding of the common sequences can not be broken during a polymerase chain reaction, resulting in no amplification of the common sequences but great amplification of the desired different sequences. The desired DNA sequences present after such covalent homologue subtraction and selective amplification represent those DNA sequences which only exist in the tester but not in the subtracter DNA library.