遗传学实验技术

The potential importance of G-quadruplex structures was implied by the recent findings that the human POT1 disrupts G-quadruplex and stimulates the telomerase activity. A solid understanding of the range of conformations that can be adopted by guanine-rich sequences can potential ...

The proximal promoter region of many human growth-related genes contains a polypurine/polypyrimidine tract that serves as multiple binding sites for Sp1 or other transcription factors. These tracts often contain a guanine-rich sequence consisting of four runs of three or more conti ...

Mounting evidence supporting the existence of DNA structures containing G-quartets in vivo makes these unique and diverse nucleic acid structures an important research subject, and future investigations aimed at elucidating their biological significance are expected. The p ...

Sequences with the potential to form intramolecular G-quadruplexes (G4-structures) are found in highly nonrandom distributions in the genomes of diverse organisms. These sequences are associated with nucleic acid metabolic processes ranging from transcription and transl ...

Methods for the molecular modeling and simulation of G-quadruplex structures and their drug/ligand complexes are discussed, and a range of protocols is presented for undertaking a variety of tasks including model-building, ligand docking, dynamics simulation, continuum solve ...

G-quartets are square planar arrangements of four guanine bases, which can form extraordinarily stable stacks when present in nucleic acid sequences. Such G-quadruplex structures were long regarded as an in vitro phenomenon, but the widespread presence of suitable sequences in geno ...

Currently, small molecules able to interact specifically with G-quadruplex DNA (G-quadruplex ligands) are intensively studied since they appear to inhibit the growth of cancer cells via an unprecedented mode of action based on structural perturbation of telomeres. It is thus of cruci ...

Telomerase inhibition through guanine quadruplex sequestration by small-molecule drugs is of great current interest as an anticancer strategy. G-quadruplexes (GQs) can be formed at the guanine-rich sequences found at the end of the telomere. They possess a large electron-rich π-sur ...

DNA can adopt a variety of non-standard conformations, including structures known as G-quadruplexes (G4-DNA), which consist of stacked tetrads of guanines. There are growing indications that G4-DNA is of biological importance, including evidence that it plays roles in telomere func ...

Verification of the existence of quadruplex structure in native human telomeres and determination of the major structure of d(T2AG3)4 (H24) in K+ solution are the major questions regarding the structure of human telomeres. We have synthesized a fluorescent probe of 3,6-bis(1-methyl-4- ...

Remarkable success in optimizing complex properties within DNA and proteins has been achieved by directed evolution. In contrast to various random mutagenesis methods and high-throughput selection methods, the number of available DNA shuffling procedures is limited, and proto ...

Differential gene expression is tightly controlled by transcription factors (TFs), which bind close to target genes and interact together to activate and coregulate transcription. Bioinformatics analysis of published genome-wide gene expression data has allowed the develo ...

Emulsion PCR, a hyper, multi, parallel PCR in water-phase droplets in water-in-oil (w/o) emulsion, can be used to make a genetic l ibrary on beads (GLOBE). In GLOBE, we have developed a novel high-throughput screening system for the analysis of the recognition sequences of DNA-binding proteins, whi ...

The RNA genomes of alphaviruses have been exploited to create highly efficient vectors for transient expression of foreign genes in mammalian cells and for use as vehicles for genetic vaccines (1–6). For Semliki Forest virus (SFV), a representative alphavirus, three strategies for in vitro ...

Inducible transgenic mouse models allow for the activation of genes in specific cells and tissues at specific times. Expression levels are dependent on the dose of the agent administered. Effective experimental models are characterized by low background levels of the regulated gene and ...

Over the past decades, transgenic and knock-out mouse models have become common use in research laboratories. Detailed phenotypic characterization of such models is essential for understanding basic mechanisms of normal physiology and disease. Hereto, pathological examina ...

Cryopreservation of mouse spermatozoa has become the foremost technique for preserving large numbers of different strains of mice with induced mutations. Recently, we have established procedures for cryopreservation of mouse spermatozoa and in vitro fertilization using cry ...

From the hybrid creatures of the Greek and Egyptian mythologies, the concept of the chimera has evolved and, in modern day biology, refers to an organism comprises of at least two populations of genetically distinct cells. Mouse chimeras have proven an invaluable tool for the generation of genet ...

Introduction of germ line mutations in mice via genetic engineering involves alterations of the structure and characteristics of genes. These alterations are mostly introduced via molecular genetic technology either in embryonal stem cells or in one-cell stage embryos. This chap ...

Recombinant adenoviruses are attractive vectors for short-term expression in mouse liver and primary cell lines. Various versatile vector systems have been developed which can be used for the reliable production of recombinant adenoviruses. This protocol describes the entire p ...