共查询到20条相似文献,搜索用时 14 毫秒
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Knox DP 《International journal for parasitology》2004,34(2):139-152
Molecular biology has provided the means to identify parasite proteins, to define their function, patterns of expression and the means to produce them in quantity for subsequent functional analyses. Whole genome and expressed sequence tag programmes, and the parallel development of powerful bioinformatics tools, allow the execution of genome-wide between stage or species comparisons and meaningful gene-expression profiling. The latter can be undertaken with several new technologies such as DNA microarray and serial analysis of gene expression. Proteome analysis has come to the fore in recent years providing a crucial link between the gene and its protein product. RNA interference and ballistic gene transfer are exciting developments which can provide the means to precisely define the function of individual genes and, of importance in devising novel parasite control strategies, the effect that gene knockdown will have on parasite survival. 相似文献
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The development of efficient methods to transfer genes into eukaryotic cells is important for molecular biotechnology. A number of different technologies to mediate gene transfer have been developed over the last 35 years, but most have drawbacks such as cytotoxicity, low efficiency and/or restricted applicability. Activated polyamidoamine (PAMAM)-dendrimers provide a new technology for gene transfer that offers significant advantages over classical methods. Reagents based on this technology provide high gene transfer efficiencies, minimal cytotoxicity, and can be used with a broad range of cell types. This technology could also be useful for in vivo gene transfer in gene therapy applications. 相似文献
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Robl JM 《Theriogenology》1999,51(2):499-508
Mammalian cloning technologies originally developed as methods of testing hypotheses about the mechanisms of cell differentiation. Embryo splitting procedures demonstrated that each of the cells in the early embryo are capable of developing into a complete new individual. Nuclear transplantation technologies have shown that loss of genetic sequences or even irreversible repression of gene function are also not mechanisms of cell differentiation. Therefore, both of these methods can be used for producing genetically identical animals. Nuclear transplantation has the advantage of being able to produce unlimited numbers of identical offspring. Highly efficient procedures have been developed for nuclear transplantation in mammals and several important characteristics of donor cells have been described. Unfortunately, the efficiency of producing cloned offspring is still low and many factors affecting the development of nuclear transfer embryos to term remain to be investigated. The tremendous potential of the technology for use in agriculture and medicine, however, will ensure that these problems are addressed and solved. 相似文献
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Ex vivo gene transfer and correction for cell-based therapies 总被引:1,自引:0,他引:1
Naldini L 《Nature reviews. Genetics》2011,12(5):301-315
Cell-based therapies are fast-growing forms of personalized medicine that make use of the steady advances in stem cell manipulation and gene transfer technologies. In this Review, I highlight the latest developments and the crucial challenges for this field, with an emphasis on haematopoietic stem cell gene therapy, which is taken as a representative example given its advanced clinical translation. New technologies for gene correction and targeted integration promise to overcome some of the main hurdles that have long prevented progress in this field. As these approaches marry with our growing capacity for genetic reprogramming of mammalian cells, they may fulfil the promise of safe and effective therapies for currently untreatable diseases. 相似文献
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Parker AL Newman C Briggs S Seymour L Sheridan PJ 《Expert reviews in molecular medicine》2003,5(22):1-15
Medical research continues to illuminate the origins of many human diseases. Gene therapy has been widely proposed as a novel strategy by which this knowledge can be used to deliver new and improved therapies. Viral gene transfer is relatively efficient but there are concerns relating to the use of viral vectors in humans. Conversely, nonviral vectors appear safe but inefficient. Therefore, the development of an efficient nonviral vector remains a highly desirable goal. This review focuses on the numerous challenges preventing efficient nonviral gene transfer in vivo and discusses the many technologies that have been adopted to overcome these problems. 相似文献
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《TARGETS》2003,2(6):253-260
Inhibition of gene expression using the RNA interference (RNAi) pathway is rapidly becoming the method of choice for studying gene function in mammalian cells. However, successful knockdown of the target gene requires efficient delivery of short interfering RNAs (siRNAs). Several technologies have been developed that enable effective delivery of siRNAs to both cells in culture and whole animals. These technologies will allow the use of RNAi to study gene function in mammalian model systems in which classical methods are often limited and costly. 相似文献
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Direct DNA transfer methods based on particle bombardment have revolutionized plant genetic engineering. Major agronomic crops
previously considered recalcitrant to gene transfer have been engineered using variations of this technology. In many cases
variety-independent and efficient transformation methods have been developed enabling application of molecular biology techniques
to crop improvement. The focus of this article is the development and performance of electric discharge particle bombardment
(ACCELL™) technology. Unique advantages of this methodology compared to alternative propulsion technologies are discussed
in terms of the range of species and genotypes that have been engineered, and the high transformation frequencies for major
agronomic crops that enabled the technology to move from the R&D phase to commercialization.
Creation of transgenic soybeans, cotton, and rice will be used as examples to illustrate the development of variety-independent
and efficient gene transfer methods for most of the major agronomic crops. To our knowledge, no other gene transfer method
based on particle bombardment has resulted in variety-independent and practical generation of large numbers of independently-derived
crop plants. ACCELL™ technology is currently being utilized for the routine transfer of valuable genes into elite germplasm
of soybean, cotton, bean, rice, corn, peanut and woody species. 相似文献
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The concept that the outcome of a devastating disease can be modified by inserting a transgene into abnormal cells is appealing. However, the gene-transfer technologies that are available at present have limited the success of gene therapy so far. Nevertheless, severe combined immunodeficiencies are a useful model, because gene transfer can confer a selective advantage to transduced cells. In this way, a proof of concept for gene therapy has been provided. 相似文献
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High-throughput proteomics: protein expression and purification in the postgenomic world. 总被引:6,自引:0,他引:6
S A Lesley 《Protein expression and purification》2001,22(2):159-164
Proteomics has become a major focus as researchers attempt to understand the vast amount of genomic information. Protein complexity makes identifying and understanding gene function inherently difficult. The challenge of studying proteins in a global way is driving the development of new technologies for systematic and comprehensive analysis of protein structure and function. Protein expression and purification are key processes in these studies, but have typically only been applied on a case-by-case basis to proteins of interest. Researchers are addressing the challenge of parallel expression and purification of large numbers of gene products through the principles of high-throughput screening technologies commonly used in pharmaceutical development. Some of the issues relevant to these approaches are discussed here. 相似文献
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The success of hematopoietic stem cell gene therapy for X-linked severe combined immunodeficiency (X-SCID) was a major breakthrough in the field of gene therapy. However, two patients treated with this gene therapy developed leukemia at a later time, and retroviral vector-mediated gene transfer was considered to trigger leukemogenesis; i.e. insertional mutagenesis caused activation of LMO 2 gene, which was one step toward leukemia development. To cope with this serious problem, basic studies are required to improve the safety of retroviral vectors and to develop the method for site-specific integration of transgenes. In addition, we have to develop technologies such as selective amplifier genes (SAGs), the system for selective expansion of transduced cells, in order to obtain therapeutic efficacy of hematopoietic stem cell gene therapy in many other disorders. Moreover, clinical applications of AAV vector are promising from the standpoint of safety issue, because this vector is derived from non-pathogenic virus. AAV vector is appropriate for gene transfer into neurons, muscles, and hepatocytes. For example, gene therapy for Parkinson's disease is investigated using AAV vectors. Genetic manipulation is also one of the indispensable technologies in the field of regeneration medicine, and further promotion of basic research is important. 相似文献
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The genetic and physiological similarities between mice and humans have focused considerable attention on rodents as potential
models of human health and disease. Together with the wealth of resources, knowledge, and technologies surrounding the mouse
as a model system, these similarities have propelled this species to the forefront of biomedical research. The advent of genomic
manipulation has quickly led to the creation and use of genetically engineered mice as powerful tools for cutting edge studies
of human disease research including the discovery, refinement, and utility of many currently available therapeutic regimes.
In particular, the creation of genetically modified mice as models of human disease has remarkably changed our ability to
understand the molecular mechanisms and cellular pathways underlying disease states. Moreover, the mouse models resulting
from gene transfer technologies have been important components correlating an individual’s gene expression profile to the
development of disease pathologies. The objective of this review is to provide physician-scientists with an expansive historical
and logistical overview of the creation of mouse models of human disease through gene transfer technologies. Our expectation
is that this will facilitate on-going disease research studies and may initiate new areas of translational research leading
to enhanced patient care. 相似文献
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Systematic approaches to mouse mutagenesis 总被引:13,自引:0,他引:13
A major challenge in post-genomics is the systematic determination of mammalian gene function. A variety of mouse mutagenesis technologies, both gene- and phenotype-driven, are being used to underpin systematic and comprehensive approaches to mammalian gene function studies. Recently, a number of centres have completed large-scale ENU mutagenesis programmes that employ a phenotype-driven approach to the generation of mouse mutants. The use of ENU mutagenesis represents a powerful and efficient approach to mammalian gene-function studies, but many parallel developments are needed in downstream technologies to properly harness the new enlarged mouse-mutant resources that are being created. 相似文献
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The transfer of naked deoxyribonucleic acid (DNA) represents an alternative to viral and liposomal gene transfer technologies
for gene therapy applications. Various procedures are employed to deliver naked DNA into the desired cells or tissues in vitro
and in vivo, such as by simple needle injection, particle bombardment, in vivo electroporation or jet injection. Among the
various nonviral gene delivery technologies jet injection is gaining increasing acceptance because it allows gene transfer
into different tissues with deeper penetration of the applied naked DNA. The versatile hand-held Swiss jet injector uses pressurized
air to force small volumes of 3 to 10 μL of naked DNA into targeted tissues. The β-galactosidase (LacZ) reporter gene construct and tumor necrosis factor α gene-expressing vectors were successfully jet injected at a pressure
of 3.0 bar into xenotransplanted human tumor models of colon carcinoma. Qualitative and quantitative expression analysis of
jet injected tumor tissues revealed the efficient expression of these genes in the tumors. Using this Swiss jet-injector prototype
repeated jet injections of low volumes (3–10 μL) into one target tissue can easily be performed. The key parameters of in
vivo jet injection such as jet injection volume, pressure, jet penetration into the tumor tissue, DNA stability have been
defined for optimized nonviral gene therapy. These studies demonstrate the applicability of the jet injection technology for
the efficient and simultaneous in vivo gene transfer of two different plasmid DNAs into tumors. It can be employed for nonviral
gene therapy of cancer using minimal amounts of naked DNA. 相似文献
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High-efficiency gene transfer into cultured embryonic motoneurons using recombinant lentiviruses 总被引:1,自引:1,他引:0
Bender FL Fischer M Funk N Orel N Rethwilm A Sendtner M 《Histochemistry and cell biology》2007,127(4):439-448
Primary neurons are a common tool for investigating gene function for survival and morphological and functional differentiation.
Gene transfer techniques play an important role in this context. However, the efficacy of conventional gene transfer techniques,
in particular for primary motoneurons is low so that it is not possible to distinguish whether the observed effects are representative
for all neurons or only for the small subpopulation that expresses the transfected cDNA. In order to develop techniques that
allow high gene transfer rates, we have optimized lentiviral-based gene transfer for cultured motoneurons by using a replication-defective
viral vector system. These techniques result in transduction efficacies higher than 50%, as judged by EGFP expression under
the control of SFFV or CMV promoters. Under the same conditions, survival and morphology of the cultured motoneurons was not
altered, at least not when virus titers did not exceed a multiplicity of infection of 100. Under the same cell culture conditions,
electroporation resulted in less than 5% transfected motoneurons and reduced survival. Therefore we consider this lentivirus-based
gene transfer protocol as a suitable tool to study the effects of gene transfer on motoneuron survival, differentiation and
function. 相似文献
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Paul Christou 《In vitro cellular & developmental biology. Plant》1993,29(3):119-124
Summary Commercialization and marketing of agricultural products derived from recombinant DNA technologies is now becoming a reality.
Genetically engineered tomatoes are poised to appear in supermarket shelves in the United States sometime next year, with
other crops, including cotton, maize, soybean, rapeseed, potatoes, cucurbids, raspberries and others, scheduled for release
1 to 2 yr later (Hamilton and Ellis, 1992). One of the major breakthroughs responsible for the rapid creation of commercializable
products has been the development of gene transfer methods capable of introducing foreign DNA into elite germplasm. This review
examines some of the principles and applications of such gene transfer technologies in relation to conventional alternatives
that are limited by cell culture, host, and genotype. Advantages and disadvantages of various gene transfer methods will be
discussed. Special emphasis will be given to particle bombardment, as methods based on this technology paved the way for the
engineering of a number of important agronomic species.
Presented in the Session-in-Depth New Approaches to Plant transformation at the 1992 World Congress on Cell and Tissue Culture,
Washington, DC, June 20–25, 1992. 相似文献