遗传学实验技术

RNA interference (RNAi) is one of the most commonly used techniques for examining the function of genes of interest. In this chapter we present two examples of RNAi that use the particle inflow gun for delivery of the DNA constructs. In one example transient RNAi is used to show the function of an anthocya ...

The ability to generate transgenic animals to study gene expression and function is a powerful and important part of the Caenorhabditis elegans genetic toolbox. Transgenic animals can be created by introducing exogenous DNA into the worm germline either by microinjection or by microp ...

The generation of transgenic animals is an essential part of research in Caenorhabditis elegans. One technique for the generation of these animals is biolistic bombardment involving the use of DNA-coated microparticles. To facilitate the identification of transgenic animals wi ...

Biolistics has become a versatile tool for direct gene transfer to various cell and tissue types. Following its successful use on the parasitic nematode Ascaris suum, we developed and evaluated biolistics in the transfection of the model filarial parasite Brugia malayi. Biolistics was p ...

The development of cervical cancer is associated with infection by oncogenic human papillomaviruses (HPVs), of which type 16 (HPV16) is the most prevalent in HPV-induced malignant diseases. The viral oncoproteins E6 and E7 are convenient targets for anti-tumor immunization. To adapt t ...

DNA immunization by gene gun against a variety of infectious diseases has yielded promising results in animal models. Skin-based DNA vaccination against these diseases is not only an attractive option for the clinic but can aid in the discovery and optimization of vaccine candidates. Vacc ...

The hepatitis B and C viruses (HBV/HCV) are major causes for chronic liver disease globally. For HBV new antiviral compounds can suppress the viral replication for years, but off-therapy responses are rare. Current therapies based on interferon and ribavirin can cure 45–85% of the treated HCV- ...

Particle-mediated DNA delivery technologies (“gene guns”) have been shown in both animal and clinical studies to be an effective means of increasing the immunogenicity and protective efficacy of DNA vaccines. The primary goal in optimizing particle-mediated epidermal delivery (P ...

Gene gun administration of DNA is an invaluable technique for transfecting tissues with only 1 μg DNA/shot. Here, we describe a transfection technique of healthy corneas of BALB/c mice with a standard gene gun, using a technique that can avoid tissue destruction even when high pressure is used (e.g., ...

The transcriptional targeting of gene expression to selected cells by cell type-specific promoters displays a fundamental tool in gene therapy. In immunotherapy, dendritic cells (DCs) are pivotal for the elicitation of antigen-specific immune responses following gene gun-med ...

Because they specifically kill dividing cells, untargeted chemotherapeutic drugs such as platin derivatives, antimetabolites or topoisomerase inhibitors for example impact the immune system resulting in more or less profound transient lympho- and/or myelo-ablations in t ...

Chemokines are a family of small, secreted proteins that function in leukocyte and tumor cell trafficking and recruiting. CC chemokine ligand 21 (CCL21)/secondary lymphoid chemokine (SLC) belongs to the inflammatory subgroup of chemokines and is expressed by stromal cells in the T-cel ...

Biolistic DNA delivery is an approach to deliver plasmid to culture cells, plants, or animals. Plasmid DNA is usually transferred through bombardment of DNA-coated particles by highly pressurized gas in various kinds of delivery vehicles. The low pressure gene gun can deliver plasmid at lo ...

Transfection of postmitotic neurons is one of the most challenging goals in the field of gene delivery. Currently most procedures use dissociated cell cultures but organotypic slice preparations have significant advantages as an experimental system; they preserve the three-dim ...

Transfection of mammalian cells has long been an extremely powerful approach for the study of the effects of specific gene expression on cell function. Until recently, however, this approach has been unavailable for the study of gene function in adult cardiac myocytes. Here, an adaptation of t ...

A nonviral method for gene transfer into mammalian cells has been developed using physical force which accelerates plasmid DNA-coated gold particles to high -speed and penetrate the mammalian cells. This technology of gene transfer via a biolistic transfection method has been shown to h ...

Tuberculosis (TB) has been listed as one of the most prevalent and serious infectious diseases worldwide. The etiological pathogen of TB is Mycobacterium tuberculosis (Mtb), a facultative intracellular bacterium. Mycobacterium bovis bacillus Calmette-Gu�rin (BCG) is the only ap ...

Immunization to protect against Trypanosoma cruzi infection has the potential to greatly decrease the burden of Chagas’ disease in the Americas. Several target antigens have been explored by multiple investigators and show promise, but given that this parasite has multiple stages wi ...

We describe here the use of particle-mediated gene transfer for the induction of immune responses against melanoma antigens in murine tumor models using the melanocyte differentiation antigen tyrosinase-related protein 2 (TRP2) as an antigen in a murine B16 melanoma model. We have util ...

DNA-based vector systems have been widely studied as new modalities for the prevention and treatment of human diseases. As for all other medicinal products, safety is an important aspect in the evaluation of such products. In this chapter we reflect on the basic safety issues which have been raised ...