Small interefence RNA (siRNA) has revamped the technology of gene silencing in cultured mammalian cells after its first demonstration by Tushl et al. 4 yr ago. To circumvent the cost and the inconvenience in identifying a unique siRNA duplex that can quench target gene expression, we devised a rep ...

MicroRNAs (miRNAs) have a vital role in the generation of plant forms through posttranscriptional regulation of the accumulation of developmental regulators. Analysis of their roles requires detailed knowledge of their expression patterns. We describe an in situ hybridization ...

MicroRNAs (miRNAs) are ubiquitous regulators of gene expression in plants and ani-mals. Their distinctive structure, as very short RNAs with a 5′-phosphate and 3′-hydroxyl group, has enabled the development of protocols to clone miRNAs. After enrichment of these small molecules by size, s ...

MicroRNAs (miRNAs), an abundant class of approx 22-nucleotide (nt) small RNAs that control gene expression at the posttranscriptional level, may play important roles during normal hematopoiesis and leukemogenesis. This chapter focuses on the methods and strategies for dissecti ...

Animal genomes contain on the order of at least hundreds of microRNAs (miRNAs). Although most remain uncharacterized, it is already clear that miRNAs regulate many bio-logical processes. A number of Drosophila miRNAs have been identified as likely cell death regulators, but functions for ...

Initially reported for Caenorhabditis elegans, microRNA (miRNA) has been shown to regulate gene expression in plants, flies, and mammals (1,2). Recently, findings of miRNA have been extended to viruses (3,4). Here, we report on our approaches to investigate the role of miRNAs in human immunode ...

MicroRNAs (miRNAs) are 21- to 25-nucleotides (nt) long and interact with messenger RNAs to trigger either translational repression or RNA cleavage through RNA interference (RNAi). We have shown that HIV-1 nef double-stranded RNA from AIDS patients who are long-term nonprogressors, inh ...

Fragile X syndrome, a common form of inherited mental retardation, is caused by the loss of the Fragile X mental retardation protein (FMRP). FMRP, which may regulate translation in neurons, not only associates with specific messenger RNAs (mRNAs) and with microRNAs (miRNAs), but also associa ...

Eukaryotic CDC6 gene function is required for the initiation of DNA replication and is a key regulatory protein during cell cycle progression. The human CDC6 gene is not expressed in most normal tissues, in contrast with its marked expression in proliferating cancer cells. An effective way to e ...

Recent studies have shown that the microRNA (miRNA) pathway is an evolutionarily conserved endogenous pathway that is important for normal development. Mature miRNAs are excised from precursors in a stepwise process and subsequently incorporated into an RNA-induced silencing c ...

MicroRNAs (miRNAs), small single-stranded regulatory RNAs capable of interfering with intracellular messenger RNAs (mRNAs) that contain either complete or partial complementarity, are useful for the design of new therapies against cancer polymorphism and viral mutation. Nu ...

Prediction of microRNA (miRNA) candidates using computer programming has identified hundreds and hundreds of genomic hairpin sequences, of which, the functions remain to be determined. Because direct transfection of hairpin-like miRNA precursors (pre)-miRNAs in mammalian c ...

As more and more genes are cloned and sequenced, it is apparent that nearly all genes are related to other genes. Similar genes are grouped into families, such as the collagen and globin gene families. There are also gene superfamilies. Gene superfamilies are composed of genes that have areas of high ho ...

The Lambda ZAP� vectors have been designed to allow in vivo excision and recircularization of the cloned insert and phagemid sequences contained within the λ vector (1). Once a Lambda ZAP library is constructed and amplified, putative clones or the λ library itself may be excised into the phagemid f ...

The starting material for any representative plant cDNA library is a supply of good-quality messenger RNA from the plant tissue of choice. Extraction of RNA can be made from several grams of tissue or as little as 50 mg. However, large samples are generally more representative of the genes expressed in a ...

It is estimated that 50,000–100,000 genes are expressed across the cell types of higher eukaryotes. As a consequence of differential expression (either regionally, temporally, or environmentally specific), a large proportion of all transcripts (approx 40–45%) represent low-abun ...

The way cDNA libraries are used for finding genes of interest depends on the overall strategy adopted. Therefore, before exploring the options for cDNA library access, we briefly delineate these strategies and the place of cDNA libraries within them.

The accumulation and analysis of expressed sequence tags (ESTs) are now an important component of genome research (1). An EST is a fragment of a sequence from a cDNA clone that corresponds to an mRNA. ESTs have applications in the discovery of new human genes, mapping of the human genome, and identificat ...

The Human cDNA Database (HCD) is a repository for human cDNA sequences and related data that is curated and maintained at The Institute for Genomic Research (TIGR). The foundation of the database is ∼160,000 partial cDNA sequences that have been generated by TIGR and Human Genome Sciences (HGS). Th ...

Since the first report on cDNA cloning in 1972 (1), this technology has been developed into a powerful and universal tool in the isolation, characterization, and analysis of both eukaryotrc and prokaryotic genes. But the conventional methods of cDNA cloning require much effort to generate a li ...