遗传学实验技术

SV40 is a small simian virus that has been extensively used as a viral vector in mammalian cells (1–4). The viral genome is a double-stranded supercoiled DNA molecule of 5243 bp (Fig. 1A) that is packaged into a capsid containing the viral structural proteins VP1,VP2, and VP3. SV40 induces a lytic infecti ...

Vaccinia virus has been used to express many diverse genes, such as prokaryotic enzymes, eukaryotic growth factors, protozoan structural proteins, and 50 different virus gene products (for reviews, seerefs. 1–3). The widespread use of vaccinia virus as a vector owes much to the ease of generat ...

Adenoviruses have been isolated from a large number of different species (mammalian and fowl) and over 100 different serotypes have been reported, some 43 of them human. The human adenoviruses, particularly types 2, 5, and 12, have been the most extensively characterized, and these viruses have ...

McBride and Ozer (1) were the first to show that purified metaphase chromosomes could act as vectors in transferring genetic information into mammalian cells. This technique, termed chromosome-mediated gene transfer (CMGT), involves the transfer of subchromosomal fragments from ...

Type 1 herpes simplex virus (HSV-1)-based vectors, which are naturally capable of carrying large DNA fragments like the 14 kb dystrophin cDNA, have been studied for their ability to transduce muscle cells (1–5). These vectors can persist in the host cell in a nonintegrated state and can be prepared at a ...

Gene transfer to the nervous system is an attractive option to treat a wide variety of neurological insults (1–3). The expression of trophic factor and/or antiapoptotic genes may be beneficial in halting the slow neurodegeneration in such conditions as Parkinson’s disease (4,5), the rapid ne ...

Glial tumors occur as intraaxial masses in the brain and are uniformly fatal due to lack of effective therapy. Resection combined with radiation and chemotherapy fails to eradicate malignant cells infiltrating into normal brain, and recurrence at the original site is ultimately fatal. G ...

In contrast to traditional drugs that generally act by altering existing gene product function, gene therapy aims to target the root cause of the disease by altering the genetic makeup of the cell to treat the disease. Researchers have adapted several classes of viruses as gene-transfer vecto ...

A relatively recent advance in the use of recombinant baculoviruses is their use for delivery of genes and genetic elements into mammalian cells. Baculovirus vectors retrofitted with mammalian gene promoters have been shown to efficiently deliver and express genes in a broad assortme ...

For more than two decades, retroviral biology has been the most intensely studied field in virology. The retroviral genome is encoded by a 7–11 kb positivesense single-stranded RNA molecule, two of which homodimerize and package in lipid-enveloped viral particles. Following attachme ...

Vectors derived from oncoretroviruses, represented by the prototype Molo-ney murine leukemia virus (MMLV), are powerful tools for gene transfer into mammalian cells. Vectors derived from such viruses are able to carry an insert of up to 6.5 kb. Because Retroviridae and derived vectors ins ...

Hematopoietic stem cells (HSCs) are clonogenic cells capable of both self-renewal and multilineage differentiation. An efficient method for gene transfer into HSCs is required for exploring HSC biology as well as for gene therapy of hematopoietic disorders. Retroviral vectors have ...

Recombinant viral vectors have been used to study a variety of fundamental issues in developmental neurobiology, as well as pathogenesis and treatments for various neurodegenerative diseases. Lentiviral vectors are valuable tools for neurobiology research owing to their abil ...

The primary aim of gene transfer into the retinal cells has been to investigate the developmental mechanisms of the retinal cells or to reverse retinal diseases. Retroviruses have been used to investigate the differentiation of retinal cells, to study the embryonic retina in vivo or explant o ...

Dendritic cells (DCs) are potent antigen-presenting cells (APCs) that play a pivotal role in stimulating antigen-specific T cells in vivo (1,2). The cardinal properties of DCs are: (1) the ability to take up, process, and present antigens; (2) the ability to migrate through different tissues into ...

Viruses have evolved to deliver their genetic cargo to cells and, due to the pathogenicity of some viruses, this process has been the subject of a great deal of study. In this respect, retroviruses came to the fore in the early 1900s with the demonstration by Ellermann and Bang (1908) and by Rous (1911) that chi ...

Moloney leukemia virus-based vectors can be generated in cells that express the products of three retroviral genes, gag, pol, and env. There are a number of cell lines such as PG13 (1) and FLYA13 (2), known as packaging cells, that have been established that stably express these genes. When these cells are t ...

Vectors derived from retroviruses have been widely studied as tools for gene transfer into mammalian tissue in vivo. One application for which retroviral vectors have received particular attention is gene transfer into tumor cells for treatment of cancer. Simple retroviruses, such as ...

Bone marrow hematopoiesis is maintained by hematopoietic stem cells (HSC) (1). Because of their unique features to self-renew and differentiate along all lineages of hematopoietic cells, even a single HSC can completely reconstitute bone marrow hematopoiesis of irradiated recip ...

 With the completion of the mouse genome sequence an essential task for biomedical sciences in the twenty-first century will be the generation and functional analysis of mouse models for every gene in the mammalian genome. More than 30,000 mutations in ES cells will be engineered and thousands of ...