细胞技术

Antisense-mediated therapy holds great promise for the treatment of central nervous system (CNS) diseases in which neurodegeneration is linked to over-production of endogenous protein. Administration of antisense therapy could be difficult, however, because peripherally ...

The development of new anticancer drugs and the identification of novel targets are a major focus for pharmaceutical and biotech companies, universities, and research institutes worldwide (1). However, the therapeutic efficacy of anticancer drugs against malignant diseases is l ...

Radiation and traditional chemotherapy kill both normal and tumor cells owing to lack of target selectivity. In this chapter, we review methods for the use of antisense oligodeoxynucleotides (AS-DONs) directed against gene products overexpressed in tumor cells in synergy with other a ...

Various genetic abnormalities accumulate during multistage carcinogenesis. The changes include structural aberrations such as point mutation, amplification, and deletion, as well as functional alterations such as abnormal level and timing of gene expression (1,2). Althou ...

Cancer of the exocrine pancreas will attack an estimated 31,000 Americans in 2004, causing an estimated 30,000 deaths (1). Cancer of the exocrine pancreas (hereafter simply referred to as pancreatic cancer) occurs more frequently in African-Americans than in European-Americans. The d ...

Nucleic acid molecules have emerged as versatile tools with promising utility as therapeutics for human diseases. The specificity of hybridization of an antisense oligonucleotide (AS-ODN) to the target mRNA makes the antisense strategy attractive for selectively modulating t ...

Until now, the clinical demise of cancer has relied on surgical resection and the inhibition of tumor cell proliferation using ionizing radiation or chemotherapeutic drugs designed to perturb DNA synthesis or the mitotic event. The development of cytotoxic agents has resulted in impro ...

In the mammalian heart, intracellular Ca2+ movements are tightly regulated at various levels within the cell (1). The sarcoplasmic reticulum (SR) plays an important role in orchestrating the movement of calcium during each contraction and relaxation. Excitation leads to the opening of v ...

Antisense therapeutics can be applied to hypertension. Hypertension is a widespread disease, yet in spite of several excellent drugs for the treatment of hypertension, the number of patients with controlled hypertension is about 27% in the United States. In all other countries studied, ev ...

In 1977, percutaneous transluminal coronary angioplasty (PTCA) was first introduced (1), and it has become an effective treatment for limited coronary artery disease (2). PTCA treatment has since become more extensive and gained favor as an alternative treatment for coronary artery by ...

Over a span of more than two decades, antisense strategies for gene therapy have expanded from antisense oligonucleotides (AS-ODNs) solely, to the addition of ribozymes and, more recently, to the inclusion of small interfering RNAs (siRNAs). Antisense therapeutics has also experienced ...

During the past decade, only one antisense-based therapy has received full Food and Drug Administration (FDA) approval. Vitravene™, developed by Isis Pharmaceuticals, was the first drug based on antisense technology to be successfully commercialized and used in treatment (1). The the ...

In this chapter we provide an introduction to statistical methods appropriate in G-protein-coupled receptor research, including examples. Topics covered include the choice of appropriate averages and measures of dispersion to summarize data sets, and the choice of tests of signif ...

Accumulating examples have demonstrated that knock-out and knock-in mice of G-protein-coupled receptors (GPCRs) are useful in elucidating physiological functions of the receptor in vivo. GPCR knock-out and knock-in are achieved by either (1) manipulation of the endogenous locus of ...

Numerous recent studies have suggested that the predicted cytosolic domains of G-protein-coupled receptors (GPCRs) represent a surface for association with proteins that may serve multiple roles in receptor localization, turnover, and signaling beyond the well-characteri ...

Many protein interactions with G-protein-coupled receptors (GPCRs) appear to influence receptor signaling and functional regulation. There is great interest therefore in methods for the identification of novel or unanticipated GPCR binding proteins. A proven method for iden ...

The versatility, sensitivity, and feasibility of fluorescence methods are very attractive to study protein-protein interaction at low levels of protein expression. However, one of the most severe limits in protein chemistry has been the difficulty of introducing site-specific fl ...

Complex networks of protein-protein interactions are key determinants of cellular function, including those regulated by G-protein-coupled receptors (GPCRs). Formation of either stable or transitory complexes are involved in regulating all aspects of receptor function, f ...

This chapter describes three different strategies of receptor mutagenesis with their advantages, disadvantages, and limitations. Oligonucleotide-directed mutagenesis using either the Altered Sites� II in vitro mutagenesis system or the GeneTailor� site-directed mut ...

In this chapter we describe methods for detecting the ubiquitination state of G-protein-coupled receptors (GPCRs). This involves coexpression of a GPCR with an epitope-tagged ubiquitin construct in a heterologous expression system. Modification by ubiquitin of the GPCR resulti ...