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The development of recombinant DNA technology and methods for transferring recombinant genes into plants has brought about significant advances in plant.
Table of contents
- Gene Transfer and Expression Protocols: 7 (Methods in Molecular Biology)
- Plant Gene Transfer and Expression Protocols | SpringerLink
- Plant Gene Transfer and Expression Protocols
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Saenz, R. Barraza, N. Loewen, W. Teo, and E. Poeschla 7. Lentivirus Transduction of Hematopoietic Cells M. Li and J. Rossi 8. Parveen, M. Mukhtar, and R. Pomerantz 9. Josephson and D.
Gene Transfer and Expression Protocols: 7 (Methods in Molecular Biology)
Russell Park and A. Mergia Yee Verhoeyen and F. Cosset Guibinga and T. Friedmann Vignali, and D. Vignali Ross and R. Parks Palmer and P. Ng Cell and Tissue Targeting Y. Kawakami and D. Curiel Chadeuf and A. Salvetti Asokan and R. Samulski Goins, D. Krisky, J. Wechuck, D. Wolfe, S. Huang, and J.
Glorioso Bowers and H. Federoff Whitehouse Saydam, D. Glauser, and C.
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Fraefel Polyomaviruses: SV40 D. Strayer, C. Mitchell, D. Maier, and C. Nichols Gottesman Ernst, R. Grabherr, and C. Airenne, O. Laitinen, A. Lundstrom Baird Trepel, W. Design and Characterization of Gene Transfer Vectors. DNA microarrays and gene expression.
Heterologous Gene Expression in E. Regulation of Gene Expression. Vascular Disease. Molecular Biology and Gene Transfer Protocols. Cardiac Cell and Gene Transfer. Analyzing Microarray Gene Expression Data. Serial Analysis of Gene Expression. Some scientific hurdles—such as the absence of efficient delivery systems, difficulty with sustained expression, insertional mutagenesis and host immune reactions—remain formidable challenges to the field Kay, Some practical limitations associated with even the most successful gene transfer techniques remain to be resolved before any gene transfer procedure can be demonstrated to be a safe and effective therapy Grigsby and Leong, Many of the major hurdles have to do with providing efficient gene delivery.
First, the vector uptake and distribution must be tightly controlled so that expression of the vector-encoded gene remains within the therapeutic range—if expression is too low, the functional protein product may not be produced at a high enough concentration to effectively restore the intended biochemical pathway, and if expression is too high, the research subject may experience toxic effects. Transcription of the new genetic material must also remain stable so that the transgene is expressed as long as necessary to treat the disease. For a given patient, this could range from a limited period to life-long expression Kay, The degree to which the vector containing the corrective gene is taken up in a sufficient number of target cells is influenced by vector size and stability, the extent of target tissue vasculature, and the efficiency of interactions between vector and host cell receptors.
The ideal vector would be cell-type specific, but the design of either non-viral or viral vectors that successfully target a specific cellular receptor has been elusive despite a great deal of effort. To date, re-engineered viral vectors are often too large, too unstable, or otherwise unable to reach the nucleus of some cell types Kay, Non-viral vectors are attractive because of their suitability for pharmaceutical considerations such as scale-up, storage stability, and quality control; however, non-viral gene delivery remains prohibitively inefficient for most therapeutic applications Grigsby and Leong, Second, a substantial proportion of the population has been exposed to viruses from which vectors have been derived or engineered , especially adenoviral and adeno-associated viral vectors.
Exposed individuals thus have circulating antibodies that can interfere with transduction of closely related recombinant vectors.
Third, gene transfer involves the interaction of many agents. The combined risk factors associated with the individual components, risks that may be amplified by their interaction, complicate risk assessments Kimmelman, For example, the cases of leukemia that arose in the X-linked severe combined immunodeficiency disorder trial may have been the result of the combined toxic effects of the vector and the transgene Baum et al. Finally, permanent genetic modification may expose patients to lifelong risks.
Few long-term studies have been conducted to detect potential low-level toxic side effects of gene transfer products or assess cumulative effects on patient health over time Hedman et al. The committee concluded that, although not without challenges, the field of gene transfer research has experienced dramatic advances in scientific knowledge and that somatic gene transfer clinical investigations may today be considered part of an established scientific research enterprise. Whereas the field of gene transfer research was characterized in its early years by considerable uncertainty and concern about theoretical risks—on the part of the public as well as the scientific community—this field has matured to a state in which some of the early concerns about risk, and uncertainty overall, have been minimized.
With the experience of more than 40 years of gene transfer trials and nearly 1, currently approved clinical trials; much has been learned about potential adverse events and how to ensure the safety of research participants. The committee concluded that many gene transfer clinical trials pose acceptable risks and are fast becoming an established modality of modern medicine. Although the state of gene transfer research is constantly evolving, not all of gene transfer research can still be considered a completely new scientific enterprise or novel technology.
This conclusion has significant repercussions for the oversight required for research projects to proceed; the committee's assessment of the current regulatory structure is the subject of the next chapter. Considerations of an appropriate regulatory structure, one that protects human research subjects while not adding to researchers' administrative burden unnecessarily or impeding scientific advancements, focus to a large extent on how to assess risk.
Given that such questions about risk assessment are not unique to gene transfer research—they are shared by other cutting-edge scientific and clinical research—the committee considered the question of oversight with a broader lens. In the following two chapters, the committee explores how the regulatory considerations for the evolving field of gene transfer research may shine light on potential needs in all emerging areas of clinical sciences. A Biologic License Application is a request for permission from FDA to introduce, or deliver for introduction, a biologic product into interstate commerce.
The Journal of Gene Medicine database does not present information on trials' sponsors e. Turn recording back on. National Center for Biotechnology Information , U. Search term. Understandings of Risk and Uncertainty in Initial Gene Transfer Research In the early years of gene transfer research, the members of the scientific community generally agreed that gene transfer research had new risks and contained many uncertainties. Scientific and Technical Concerns In addition to these ethical and social issues, early gene transfer research was characterized by a high degree of technical uncertainty and its associated risks.
Some Potential Hazards Found to Not Be a Problem Many original uncertainties have been replaced by scientific clarity, and fears have been alleviated by decades of experience. Technical Advances The safety profile of most viral vectors has been considerably enhanced by advances in vector design strategies as well as better understanding of molecular virology and virus—host cell relationships Vannucci et al. Clinical Research Successes A number of clinical research successes have emerged in gene transfer since Risk Assessment Even with these pivotal advances and dramatic examples of clinical success, risk assessment remains difficult in gene transfer research Deakin et al.
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Additional considerations regarding gene therapy include the following: a In vivo gene therapy can inadvertently target transgene expression to an unintended and clinically unaffected cell or tissue type, with a potential for toxicity. Scientific Hurdles Some scientific hurdles—such as the absence of efficient delivery systems, difficulty with sustained expression, insertional mutagenesis and host immune reactions—remain formidable challenges to the field Kay, Ten years of gene therapy for primary immune deficiencies.
Nonviral gene delivery: Principle, limitations, and recent progress. American Association of Pharmaceutical Scientists Journal. Side effects of retroviral gene transfer into hematopoietic stem cells. Berg P, Mertz JE. Personal reflections on the origins and emergence of recombinant DNA technology. Bersenev A, Levine BL. Convergence of gene and cell therapy. Regenerative Medicine. Immune responses to gene therapy vectors: Influence on vector function and effector mechanisms.
Gene Therapy. Lentiviral hematopoietic stem cell gene therapy benefits metachromatic leukodystrophy. Gene therapy for primary immunodeficiency. Current Opinion in Pediatrics. Lessons learned from the clinical development and market authorization of Glybera. Human Gene Therapy. Current advances and future challenges in adenoviral vector biology and targeting.
Plant Gene Transfer and Expression Protocols | SpringerLink
Current Gene Therapy. Accepting risk in clinical research: Is the gene therapy field becoming too risk-averse. Molecular Therapy. The ethics of gene therapy: Balancing the risks. Current Opinion in Molecular Therapeutics. Cancer Research. FDA U.
Food and Drug Administration. Guidance for industry: Considerations for the design of early-phase clinical trials of cellular and gene therapy products.
Plant Gene Transfer and Expression Protocols
Fredrickson DS. Friedmann T, Roblin R. Gene therapy for human genetic disease. Gene therapy clinical trials worldwide to —An update. Journal of Gene Medicine. Balancing protection and release of DNA: Tools to address a bottleneck of non-viral gene delivery. Journal of the Royal Society Interface.
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Kay MA. State-of-the-art gene-based therapies: The road ahead. Nature Reviews Genetics. Kimmelman J. Recent developments in gene transfer: Risk and ethics. British Medical Journal. Stable ethics: Enrolling non-treatment-refractory volunteers in novel gene transfer trials. The ethics of human gene transfer. An introduction to molecular medicine and gene therapy. Molecular medicine and gene therapy: An introduction. Persistence, localization, and external control of transgene expression after single injection of adeno-associated virus into injured joints.
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