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Simultaneous Fluorescence Immunophenotyping and FISH on Tumor Cells

Chromosome aberrations are regularly detected in most hematologic neoplasms and in various solid tumors and are often associated with distinct morphologic and immunophenotypic features of certain clinico-pathologic entities of tumors (1–4). Detection of these chromosome a ...

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BAC Resource for Molecular Cytogenetics

Bacterial artificial chromosomes (BACs) are ideal materials to use for the purpose of integrating DNA sequence with cytogenetic markers. They have been the major vectors used in genome sequencing. BACs are also well suited for fluorescent in situ hybridization (FISH) in that they represe ...

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Lentiviral Vectors for Gene Transfer to the Central Nervous System: Applications in Lysosomal Storage Disease Animal Models

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, because of their a ...

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A Highly Efficient Gene Delivery System Derived from Feline Immunodeficiency Virus (FIV)

Feline immunodeficiency virus (FIV) is an appealing candidate for viral vector development because recombinant FIV vectors efficiently transduce dividing and nondividing cells and thus provides an alternative to primate lentiviral vectors derived from simian immunodef ...

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A Multigene Lentiviral Vector System Based on Differential Splicing

Lentiviral vectors are promising tools for gene transfer (1–4). Like oncoret-roviral vectors, they offer the unique advantage of stably integrating into the genome of the host cell, thus providing the basis for sustained gene expression. In contrast to the classical oncoretrovirus der ...

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Production of Trans-Lentiviral Vector with Predictable Safety

Retroviruses use homologous recombination to reverse transcribe a doublestranded copy of DNA from their RNA genome. Safety advancements in retroviral vector design include the development of packaging systems that provide all of the retroviral proteins in trans to a replication- ...

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Human Immunodeficiency Virus Type 1-Based Vectors for Gene Delivery to Human Hematopoietic Stem Cells

A number of inherited and acquired disorders can potentially be treated by gene-based therapies. To be successful, gene therapy requires efficient delivery and continued expression of the therapeutic gene in the target cell. Toward this goal, a variety of methods have been developed for de ...

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Semliki Forest Viral Vectors for Gene Transfer

Semliki Forest virus (SFV), Sindbis virus, and several pathogenic encephalitis-producing viruses (e.g., Ross river virus, RRV) are members of the family Togaviridae and genus Alphavirus. A typical alphavirus is an enveloped virus with single-stranded, positive-polarity RNA geno ...

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Semliki Forest Virus (SFV) Vectors in Neurobiology and Gene Therapy

Gene delivery methods play a key role in molecular neurobiology. The possibility to efficiently deliver and express genes not only in cell lines, but also in neurons in culture in vivo has facilitated studies on the understanding of gene function. Generally, neurons are not very susceptible to g ...

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Semliki Forest Virus Vectors for Large-Scale Production of Recombinant Proteins

Recombinant protein expression is one of the basic elements of modern molecular biology. The completion of the sequencing of the human genome has, in fact, accelerated the need of rapid and efficient expression systems. Extensive efforts have been dedicated to the development of expressi ...

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Development of Foamy Virus Vectors

Retroviral vectors developed from the murine leukemia virus (MLV) were the first and most widely used vectors for gene transfer experiments (1). Delineation of their replication strategy allowed extensive manipulation of their genome and the development of vector packaging cell li ...

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Poxviral/Retroviral Chimeric Vectors Allow Cytoplasmic Production of Transducing Defective Retroviral Particles

Retroviral vectors have become important tools in gene therapy as a result of a number of highly desirable properties, including stable genomic integration, which are rarely found with other gene delivery vectors. However, after two decades of experience with this technology, some shor ...

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Construction of Multiply Disabled Herpes Simplex Viral Vectors for Gene Delivery to the Nervous System

This chapter aims to describe the main procedures involved in constructing a disabled recombinant herpes simplex virus type 1 (HSV-1) vector. However, before describing the methodology in detail, it is important to briefly consider the structure and life cycle of HSV1 and the modificatio ...

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Improved HSV-1 Amplicon Packaging System Using ICP27-Deleted, Oversized HSV-1 BAC DNA

Herpes simplex virus type 1 (HSV-1) has several unique features that make it a promising candidate for gene transfer vehicles. These include: i) a pantropic viral envelope; ii) a large genome (approx 152 kb), which confers its large transgene capacity; and iii) the ability of entering a latency state in ...

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Herpes Simplex Amplicon Vectors

The key fundamental problem for effective gene therapy remains gene delivery. Gene therapy has achieved some modest success in animal and human experiments (1–7). However, effective gene delivery remains the chief obstacle. Herpes simplex virus type 1 (HSV-1) vectors offer several impr ...

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Strategies to Adapt Adenoviral Vectors for Targeted Delivery

One of the challenges of current gene therapy vector development, concerns targeting a therapeutic gene to diseased cells with the aim of achieving sufficient gene expression in the affected tissue, while minimizing toxicity and expression in other tissues. The use of recombinant aden ...

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Use of Recombinant Adenovirus for Gene Transfer into the Rat Brain: Evaluation of Gene Transfer Efficiency, Toxicity, and Inflammatory and Immune Reac

Adenovirus (Ad) vectors have been used to successfully deliver genes into a wide variety of non-central nervous system (CNS) tissues and organs in animal models of human disease and in several human phase I clinical trials (1–3). Adenoviruses are easily purified to the high titers required for in v ...

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Generation of Adenovirus Vectors Devoid of All Viral Genes by Recombination Between Inverted Repeats

Adenovirus (Ad) vector-mediated gene transfer is useful in applications where transient, high-level transgene expression is required. In contrast to onco-retroviral vectors, Ad vectors efficiently infect nondividing cells. Most recombinant Ad vectors currently used for in v ...

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Packaging Cell Lines for Generating Replication-Defective and Gutted Adenoviral Vectors

Gene transfer using adenovirus (Ad) can be a highly efficient method for transducing cells and tissues. Wild-type Ad has a 36-kb double-stranded linear genome that encodes numerous overlapping open reading frames (1). Because wild-type Ad is infectious to humans, vectors used in the labora ...

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Improving the Transcriptional Regulation of Genes Delivered by Adenovirus Vectors

The effectiveness of gene therapy is essentially governed by the ability of the vector to reach the relevant tissue, and once there, to facilitate the expression of appropriate quantities of the gene product. Over the course of evolution, viruses have developed highly specialized and succe ...

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