遗传学实验技术

The liver represents a major target organ for gene delivery owing to its high biosynthetic capacity and access to the bloodstream. Adenoviral vectors are highly efficient gene-transfer vehicles, making them among the most promising systems for in vivo gene transfer to the liver. Following ...

In functional genomics, the use of expression libraries of DNA variants in combination with potent screening techniques is a powerful tool for gene discovery. They allow study of gene and protein function, generation of peptide variants with novel properties, as well as identification of f ...

Adenoviruses, which were first isolated in the 1950s, have been developed as gene-delivery vehicles, or vectors, since the early 1980s (1). The adenoviruses constitute the Adenoviridae family, which is divided into two genera: the Aviadenovirus genus infects only birds, whereas the Mast ...

Delivery of genes to the pulmonary vascular endothelium is a rational approach for the investigation and potential therapy of pulmonary vascular diseases. Furthermore, in view of the exposure of this vascular bed to the entire cardiac output, this technique could be used as an efficient bas ...

The majority of eukaryotic genes undergo alternative splicing, an evolutionarily conserved phenomenon, to generate functionally diverse protein isoforms from a single transcript. The fact that defective pre-mRNA splicing can generate non-functional and often toxic prote ...

The vision of using a single therapeutic agent with sufficient generality to allow application to a wide variety of diseases, yet specific enough to permit intervention at single molecular stages of the pathology, is rapidly becoming a reality through the emergence of RNA interference. RNA ...

Short interfering RNAs (siRNAs) represent new potential therapeutic tools, owing to their capacity to induce strong, sequence-specific gene silencing in cells. However, their clinical development requires new, safe, and efficient in vivo siRNA delivery methods. In this study, we rep ...

Discovery of RNA interference (RNAi) has been one of the most important findings in the last ten years. In recent years, small interfering RNA (siRNA)-mediated gene silencing is beginning to show substantial promise as a new treatment modality in preclinical studies because of its robust gene ...

RNA interference (RNAi) is an evolutionarily conserved gene-silencing mechanism in which small 19–23-nucleotide double-stranded RNA molecules, or small interfering RNAs (siRNAs), target cognate RNA for destruction with exquisite potency and selectivity. The RNAi machine ...

Malaria is a disease that kills more than a million people each year in tropical and subtropical countries. The disease is caused by Plasmodium parasites and is transmitted to humans exclusively by mosquitoes of the genus Anopheles. The lack of functional approaches has hampered study of the bi ...

Delivery is a key issue in development of clinically relevant RNAi therapeutics. Polymeric nanoparticles formed by self-assembly of polycations with siRNA can be used for extracellular delivery, cellular uptake and intracellular trafficking as a strategy to improve the therape ...

RNAi holds promise for neurodegenerative disorders caused by gain-of-function mutations. We and others have demonstrated proof-of-principle for viral-mediated RNAi in a mouse model of motor neuron disease. Lentivirus and adeno-associated virus have been used to knockdown leve ...

The occurrence of autoimmunity is strongly associated with multiple gene variants that predispose individuals to disease. The identification of the gene polymorphisms that modulate disease susceptibility is key to our understanding of disease etiology and pathogenesis. Whi ...

RNA interference is a potent gene silencing pathway initiated by short molecules of double-stranded RNA. Small interfering RNAs (siRNAs) with full sequence complementarity to mRNAs induce cleavage of their target transcripts in the cytoplasm. Recent evidence has shown, however, th ...

With the successful completion of genome sequencing projects for a variety of model organisms, the selection of candidate organisms for future sequencing efforts has been guided increasingly by a desire to enable comparative genomics. This trend has both depended on and encouraged the d ...

There are innumerable instances when one wants to manipulate cloned DNA: point mutagenesis for analysis of protein function and transcription factor binding sites, introduction of insertions to produce fusion proteins or to introduce reporter genes or motifs, and creation of delet ...

Flow cytometry and fluorescence-activated cell sorting (FACS) are techniques of great power used to screen cells rapidly for expression of particular gene products. These techniques have been of general utility in identifying and selecting populations of cells of defined charact ...

In situ polymerase chain reaction (PCR) refers to the amplification of specific nucleic acid sequences and subsequent visualization of the PCR products in tissue sections. When PCR is performed in fixed cells in suspension the term in-cell PCR is normally applied instead. For example, in-ce ...

Chromosome “painting” by fluorescent labeled probes for each human chromosome has been available since the late 1980s. The DNA or RNA radioactive isotopes have been in use since 1969 as nucleic acid probes (1,2). The introduction of fluorescent labeled probes by Bauman et al. (1980) presented an ...

Lentiviruses, such as human immunodeficiency virus (HIV), feline immunodeficiency virus (FIV), equine infectious anemia virus (EIAV), and bovine Jembrana disease virus (JDV) are members of the Retroviridae, viruses with enveloped capsids and a plus-stranded RNA genome. Like all re ...