A modular set of lacZ fusion vectors for studying gene expression in Caenorhabditis elegans, we describe a series of pla
Fire!, 1998, 48 pages, 9780590975858, Scholastic Reference, 1998 Distinct requirements for somatic and germline expression of a generally expressed Caernorhabditis elegans gene, in screening for embryonic-lethal mutations in Caernorhabditis elegans, we defined an essential gene (let-858) that encodes a nuclear protein rich in acidic and basic residues. We have named this product nucampholin. Closely homologous sequences in yeast, plants. A modular set of lacZ fusion vectors for studying gene expression in Caenorhabditis elegans, we describe a series of plasmid vectors which contain modular features particularly useful for studying gene expression in eukaryotic systems. The vectors contain the Escherichia coli β-galactosidase (βGal)-encoding region (the lacZ gene) flanked by unique polylinker. Ingestion of bacterially expressed dsRNAs can produce specific and potent genetic interference in Caenorhabditis elegans, genetic interference mediated by doublestranded RNA (RNAi) has been a valuable tool in the analysis of gene function in Caenorhabditis elegans. Here we report an efficient induction of RNAi using bacteria to deliver double-stranded RNA. This method makes. DNA transformation, publisher Summary This chapter discusses DNA transformation. DNA transformation assays in a whole organism provide experimental links between molecular structure and phenotype. Experiments with transgenic Caenorhabditis elegans start in general with. Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems, short interfering RNAs (siRNAs) are double-stranded RNAs of≈ 21-25 nucleotides that have been shown to function as key intermediaries in triggering sequence-specific RNA degradation during posttranscriptional gene silencing in plants and RNA interference. Patterns of known and novel small RNAs in human cervical cancer, recent studies suggest that knowledge of differential expression of microRNAs (miRNA) in cancer may have substantial diagnostic and prognostic value. Here, we use a direct sequencing method to characterize the profiles of miRNAs and other small RNA segments. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans, experimental introduction of RNA into cells can be used in certain biological systems to interfere with the function of an endogenous gene 1, 2. Such effects have been proposed to result from a simple antisense mechanism that depends on hybridization between. Distinct populations of primary and secondary effectors during RNAi in C. elegans, rNA interference (RNAi) is a phylogenetically widespread gene-silencing process triggered by double-stranded RNA. In plants and Caenorhabditis elegans, two distinct populations of small RNAs have been proposed to participate in RNAi:Primary siRNAs(derived from. The rde-1 gene, RNA interference, and transposon silencing in C. elegans, double-stranded (ds) RNA can induce sequence-specific inhibition of gene function in several organisms. However, both the mechanism and the physiological role of the interference process remain mysterious. In order to study the interference process, we have. Co-evolution of a broadly neutralizing HIV-1 antibody and founder virus, current human immunodeficiency virus-1 (HIV-1) vaccines elicit strain-specific neutralizing antibodies. However, cross-reactive neutralizing antibodies arise in approximately 20% of HIV-1-infected individuals, and details of their generation could provide a blueprint. Genes and mechanisms related to RNA interference regulate expression of the small temporal RNAs that control C. elegans developmental timing, rNAi is a genesilencing phenomenon triggered by double-stranded (ds) RNA and involves the generation of 21 to 26 nt RNA segments that guide mRNA destruction. In Caenorhabditis elegans, lin-4 and let-7 encode small temporal RNAs (stRNAs) of 22 nt that regulate stage. Rolling replication of short DNA circles, natural genes and proteins often contain tandemly repeated sequence motifs that dramatically increase physiological specificity and activity. Given the selective value of such repeats, it is likely that several different mechanisms have been responsible for their. Loss of the putative RNA-directed RNA polymerase RRF-3 makes C. elegans hypersensitive to RNAi, rNA interference (RNAi) is a broadly used reverse genetics method in C. elegans [1]. Unfortunately, RNAi does not inhibit all genes [2, 3]. We show that loss of function of a putative RNA-directed RNA polymerase (RdRP) of C. elegans, RRF-3, results. A high-resolution, nucleosome position map of C. elegans reveals a lack of universal sequence-dictated positioning, using the massively parallel technique of sequencing by oligonucleotide ligation and detection (SOLiD; Applied Biosystems), we have assessed the in vivo positions of more than 44 million putative nucleosome cores in the multicellular genetic model organism. Functional anatomy of a dsRNA trigger: differential requirement for the two trigger strands in RNA interference, in RNA-mediated interference (RNAi), externally provided mixtures of sense and antisense RNA trigger concerted degradation of homologous cellular RNAs. We show that RNAi requires duplex formation between the two trigger strands, that the duplex must include. Specific interference by ingested dsRNA, a genetic interference phenomenon in the nematode Caenorhabditis elegans has been described in which expression of an individual gene can be specifically reduced by microinjecting a corresponding fragment of double-stranded (ds) RNA 1. One striking feature. RNA-triggered gene silencing, double-stranded RNA (dsRNA) has recently been shown to trigger sequence-specific gene silencing in a wide variety of organisms, including nematodes, plants, trypanosomes, fruit flies and planaria; meanwhile an as yet uncharacterized RNA trigger has been shown. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract, we have developed a cell-free system for studying the synthesis of mRNA in mammalian cells. The system consists of a dialyzed and concentrated whole-cell extract derived from HeLa cells, small molecules and cofactors needed for transcription, and exogenously added. On the role of RNA amplification in dsRNA-triggered gene silencing, we have investigated the role of trigger RNA amplification during RNA interference (RNAi) in Caenorhabditis elegans. Analysis of small interfering RNAs (siRNAs) produced during RNAi in C. elegans revealed a substantial fraction that cannot derive directly from input. RNA as a target of double-stranded RNA-mediated genetic interference in Caenorhabditis elegans, introduction of exogenous double-stranded RNA (dsRNA) into Caenorhabditis elegans has been shown to specifically and potently disrupt the activity of genes containing homologous sequences. In this study we present evidence that the primary interference effects of dsRNA. by L Timmons, A Fire
