Cardiac gene promoter analysis remains an integral method in molecular cardiology and continues to provide novel insights into the transcriptional mechanisms that regulate gene expression in the myocardium. Initial studies focused on the regulated expression of contractile ...
The availability of an increasing number of vertebrate genomes has enabled comparative methods to infer functional sequences based on evolutionary constraint. Although this has proved powerful for gene identification, significant progress has also been made in uncovering gene ...
The development of biotechnology and the completion of the Human Genome Project have led to the rapid emergence of enormous amounts of genomic information. Computational resources are needed to enable researchers and clinicians to obtain quick, up-to-date information in targeted ar ...
The availability and accessibility of high-throughput and biological legacy data have allowed mathematical analyses of genome-scale metabolic networks and models. Model formulation is centered on the conservation principles of mass and charge. Thermodynamic information ...
Matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) has become a widely used technology for the detection of nucleic acids. In this chapter we introduce its use for the discovery of novel sequence polymorphisms and the identification of kn ...
Tissue specific gene knockouts using Cre recombinase can have broad applicability in murine disease models of cardiovascular disease. The Cre system has been shown to have broad experimental versatility for both temporal and spatial control of gene deletion. By and large this is achieved ...
Neonatal rat ventricular myocytes (NRVMs) cultured in vitro have been used as a model system for easily recreating and studying several cardiac molecular conditions, such as hypertrophy, oxygen deprivation, and gene expression. However, low efficiency of gene transfer has often repr ...
Lentiviruses have the capacity to enter and integrate their genetic material into cells that are not dividing. This property is retained in vectors based on these agents. They can thus effect gene delivery to cells that are difficult to transduce such as cardiac myocytes in vitro and in vivo. They are ...
This chapter describes a system using adeno-associated viral (AAV) vector to deliver hypoxia-inducible gene expression to ischemic hearts. The hypoxia induction of gene expression in this system is based on the accumulation of hypoxia-inducible factor-1 (HIF-1) in ischemic hearts a ...
The postgenomic era has been ushered in making promises for cures and functional analysis of previously unknown or uncharacterized genes. However, realization of the potential of gene therapy for neurological disorders and functional genomic analysis in the brain remains a signifi ...
The integration of foreign DNA into the genomes of host cells is of fundamental importance for viral oncology, evolution, transgenic organisms, and gene therapy applications. Expression of foreign genes in eukaryotic cells is highly dependent upon the efficiency of integration even ...
The need for safe and effective gene transfer to mammalian cells and organisms has led many investigators to focus on the defective human parvovirus, adeno-associated virus (AAV), as a gene delivery vehicle. AAV vectors have been used to deliver genes to a wide variety of mammalian cells in culture, ...
Original standard protocols for the generation of recombinant adeno-associated viruses (rAAVs) have generally involved the cotransfection of 293 cells with a rAAV plasmid vector (pAAV) and a helper/packaging plasmid, followed by over-infection with a helper virus, normally an ad ...
The adeno-associated viruses (AAVs) are dependent parvoviruses that require helper viruses for replication. They are classified based on size and structure. Because the majority of AAV isolates were first identified as contaminants of laboratory stocks of adenovirus, little is kn ...
The limited packaging capacity of adeno-associated virus (AAV) has long been recognized as one of the most troublesome obstacles for the development of this gene delivery system. This size constraint derives from the inherent properties of the wild-type AAV virus. As the smallest DNA virus, t ...
Retroviral vectors based on Moloney Murine Leukemia Virus (MLV), described more than 15 years ago (1), first entered clinical trials in 1990 (2). Since then, a greater understanding of the basic retrovirus biology and how it relates to the production of recombinant retroviral vectors has help ...
Gene expression systems that can be induced or regulated are particularly useful when the gene of interest is toxic to the cell, or when the amount of protein expressed needs to be tightly controlled, such as for gene therapy. Many systems are available that achieve high levels of induced gene expres ...
The development of retroviral vectors has provided scientists with an effective way of achieving gene transfer stable over cell generations. This is possible because integration of proviral DNA into the genome of infected cells permits duplication of the ectopic genetic informati ...
Herpes simplex virus (HSV) is an enveloped double-stranded DNA virus (see Fig. 1A-reviewed in ref. 1). The mature virion consists of the following components: Fig. 1.A. Schematic depiction of a mature HSV virion illustrating the main components of the virus particle. B. The HSV genome is organized i ...
The development of retroviral gene transfer vectors has provided an effective way to deliver foreign genes into mammalian cells. This chapter will focus on the development of a simian retrovirus (SRV) gene delivery system. This includes the identification of the minimal packaging sign ...
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