Gene therapy and regulation最新文献_第4页

LEUKEMIA STEM CELLS: STUDYING THE ROOT OF LEUKEMIA 白血病干细胞:研究白血病的根源

Gene therapy and regulation Pub Date : 2007-03-01 DOI: 10.1142/S1568558607000058

D. Pearce, D. Bonnet

{"title":"LEUKEMIA STEM CELLS: STUDYING THE ROOT OF LEUKEMIA","authors":"D. Pearce, D. Bonnet","doi":"10.1142/S1568558607000058","DOIUrl":"https://doi.org/10.1142/S1568558607000058","url":null,"abstract":"A fundamental problem in cancer research is the identification of the cell type capable of initiating and sustaining the growth of the neoplastic clone in vivo. The key to solving this problem lies on the observation made over 40 years ago that tumors are heterogeneous and thus might be maintained only by a rare subset of cells called \"cancer stem cells\" (CSCs). However, the proof of this principle was only possible after the development of modern research tools for investigating the behavior of defined cell populations in vivo. The blood-related cancer leukemia was the first disease where human CSCs, or leukemic stem cells (LSCs), were isolated. The development of quantitative xenotransplantation assays using immune-deficient mouse recipients to detect primitive human hematopoietic stem cells (HSCs) with in vivo repopulating ability and the adaptation of this model to leukemia have been instrumental. Leukemia can now be viewed as aberrant hematopoietic processes initiated by rare LSCs that have maintained or reacquired the capacity for indefinite proliferation through accumulated mutations and/or epigenetic changes. Yet, despite their critical importance, much remains to be learned about the developmental origin of LSC and the mechanisms responsible for their emergence in the course of the disease. This report will review our current knowledge on normal and leukemic stem cell development and finally demonstrate how these discoveries provide a paradigm for identification of CSCs from solid tumors. By a careful comparative analysis of the properties of CSCs and of their normal counterparts, it should be possible to pinpoint critical features amenable to efficient anti-CSC therapies.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2007-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1142/S1568558607000058","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64014749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

CELL-FREE SELECTION OF DNA-BINDING PROTEINS FOR FUTURE GENE THERAPY APPLICATIONS ∗ 未来基因治疗应用的dna结合蛋白的无细胞选择*

Gene therapy and regulation Pub Date : 2007-03-01 DOI: 10.1142/S156855860700006X

A. Sepp, F. Ghadessy, Y. Choo

{"title":"CELL-FREE SELECTION OF DNA-BINDING PROTEINS FOR FUTURE GENE THERAPY APPLICATIONS ∗","authors":"A. Sepp, F. Ghadessy, Y. Choo","doi":"10.1142/S156855860700006X","DOIUrl":"https://doi.org/10.1142/S156855860700006X","url":null,"abstract":"Engineered DNA-binding proteins, in particular zinc finger proteins (ZFPs), have broad-ranging applications in gene therapy. An engineered ZFP transcription activator targeted to the VEGF locus is currently undergoing clinical trials for the induction of angiogenesis. A number of ZFP gene switches have been developed which allow highly controllable regulation of therapeutic transgene expression based on small molecule inducers/repressors. Finally, engineered ZFP nucleases have been used to correct a gene sequence in a living cell by stimulating homologous DNA recombination, suggesting a new, highly targeted approach to gene therapy. All these approaches rely on DNA-binding protein engineering, which in the past has mainly been achieved by selection using phage display. However, a convenient cell-free selection method known as in vitro compartmentalization (IVC) has previously been used to engineer DNA-binding proteins with enzymatic activities (e.g. polymerase and methylase), and the method has recently been extended to the engineering of sequence-specific ZFP DNA-binders. Below we describe the IVC procedure and review the progress made in applying this to the problem of facilitating the engineering of DNA-binding proteins.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2007-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1142/S156855860700006X","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64014763","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Method — A nonviral gene transfer method for transfecting multipotent adult progenitor cells (MAPC) 方法-一种转染多能成体祖细胞(MAPC)的非病毒基因转移方法

Gene therapy and regulation Pub Date : 2004-12-01 DOI: 10.1163/1568558043967481

U. Lakshmipathy, Luke Hammer, C. Verfaillie

{"title":"Method — A nonviral gene transfer method for transfecting multipotent adult progenitor cells (MAPC)","authors":"U. Lakshmipathy, Luke Hammer, C. Verfaillie","doi":"10.1163/1568558043967481","DOIUrl":"https://doi.org/10.1163/1568558043967481","url":null,"abstract":"Multipotent adult progenitor cells (MAPC) are stem cells isolated primarily from adult bone marrow that have the ability to differentiate in vitro into cells with phenotypic and functional characteristics of cells from the three germ layers, namely endoderm, mesoderm and neuroectoderm. In addition, MAPCs proliferate for extended periods of time without obvious senescence. Hence, MAPCs may be ideal cells for therapy of genetic disorders, provided that the main impediment to gene therapy, namely efficient gene transfer and persistent gene expression, can be overcome. Most commercially available lipid-based methods that very highly efficiently transfect cell lines and primary cells, fail to transfect MAPC. However, 10–15% transfection of MAPC can be achieved using Superfect. However, this approach requires high density culture of MAPCs which subsequently differentiate. Between 12 and 15% transfection is achieved in MAPCs using electroporation, but this is highly toxic to MAPCs. Here, using transient expression of an enhanced green fluorescent protein (EGFP) gene, we report that nucleofection results in a transfection efficiency of over 25% with mouse MAPC without perturbing the conditions suitable for MAPC growth and maintenance, and with significantly less toxicity than electroporation. Similar results were seen for rat and human MAPC. Efficient transfection of MAPC by nucleofection offers an appealing non-viral mode of gene delivery and can be used to overexpress genes of interest in these cells. In addition, it should easily accomodate emerging site-specific integration technology, thereby culminating eventually in MAPC-mediated long-term gene therapy for genetic disorders.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/1568558043967481","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64796194","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Small RNA-mediated chromatin modification and transcriptional gene silencing 小rna介导的染色质修饰和转录基因沉默

Gene therapy and regulation Pub Date : 2004-12-01 DOI: 10.1163/1568558043967463

H. Kawasaki, Y. Fukuda, K. Taira

引用次数: 1

Transmitochondrial stem cells: from mouse models of mtDNA diseases to autologous cybrid stem cell gene therapy 线粒体干细胞:从mtDNA疾病小鼠模型到自体杂交干细胞基因治疗

Gene therapy and regulation Pub Date : 2004-12-01 DOI: 10.1163/1568558043967508

R. Bertolotti

引用次数: 2

Antisense derivatives of U7 and other small nuclear RNAs as tools to modify pre-mRNA splicing patterns U7和其他小核rna的反义衍生物作为修饰前mrna剪接模式的工具

Gene therapy and regulation Pub Date : 2004-12-01 DOI: 10.1163/1568558043967472

M. Asparuhova, R. Kole, D. Schümperli

{"title":"Antisense derivatives of U7 and other small nuclear RNAs as tools to modify pre-mRNA splicing patterns","authors":"M. Asparuhova, R. Kole, D. Schümperli","doi":"10.1163/1568558043967472","DOIUrl":"https://doi.org/10.1163/1568558043967472","url":null,"abstract":"The importance of alternative splicing for the diversity of the proteome and the large number of genetic diseases that are due to splicing defects call for methods to modulate alternative splicing decisions. Although splicing can be modulated by antisense oligonucleotides, this approach is confronted with problems of efficient delivery and the need for repeated administrations of large amounts of the oligonucleotides. Therefore we have developed methods allowing us to modulate splicing with the help of modified derivatives of the U7 small nuclear RNA involved in histone RNA 3′ end processing. Its nuclear accumulation as a stable ribonucleoprotein particle makes U7 snRNA especially useful for this purpose. In particular, U7 derivatives containing two tandem antisense sequences directed against targets upstream and downstream of an exon can induce the efficient and specific skipping of that exon. U7 expression cassettes have been successfully introduced into a great number of cell lines, primary cells or tissues with the help of lentiviral and adeno-associated viral vectors. Examples of these therapeutic strategies in the fields of β-thalassemia, Duchenne muscular dystrophy and HIV/AIDS are discussed.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/1568558043967472","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64796181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 11

Xenomitochondrial embryonic stem cells and mice: modeling human mitochondrial biology and disease 异种线粒体胚胎干细胞和小鼠:模拟人类线粒体生物学和疾病

Gene therapy and regulation Pub Date : 2004-12-01 DOI: 10.1163/1568558043967454

M. Cannon, C. Pinkert, I. Trounce

{"title":"Xenomitochondrial embryonic stem cells and mice: modeling human mitochondrial biology and disease","authors":"M. Cannon, C. Pinkert, I. Trounce","doi":"10.1163/1568558043967454","DOIUrl":"https://doi.org/10.1163/1568558043967454","url":null,"abstract":"The characterization of mitochondrial diseases has proceeded rapidly since the first descriptions of mitochondrial DNA (mtDNA)-linked disease mutations appeared in the late 1980s. To elucidate mechanisms of a variety of mitochondrial disorders and disease, both in vitro and in vivo modeling systems have been exploited. To produce these models, numerous approaches have been undertaken due to the difficulty associated with targeted mutagenesis and directed modification of the mitochondrial genome. Currently available models of mitochondrial disease are discussed in this paper, including our xenomitochondrial mice. In this model, mitochondria from one donor species are transferred to another. By doing so, cells and animals were generated with varying levels of heteroplasmy (or homoplasmy) for the introduced mitochondrial genomes. This caused graded variations in electron transport chain (ETC) dysfunction which were dependent upon the evolutionary divergence between donor and recipient. The protocol in generating these models involved the utilization of rhodamine-6G (R6G) to remove or eliminate endogenous mtDNA from the recipient cells. This paper will highlight the process and the implications of R6G treatment of mouse embryonic stem (ES) cells to create transmitochondrial cybrids. We summarize the history and mechanism of action of R6G as well as the future prospects for xenomitochondrial models toward increasing our understanding of mitochondrial biology and the dynamic interplay in signaling between mitochondria and the nucleus.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/1568558043967454","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

Digital RNA regulation of complex organisms 复杂生物体的数字RNA调控

Gene therapy and regulation Pub Date : 2004-12-01 DOI: 10.1163/1568558043967517

J. Mattick

引用次数: 0

Safe delivery of therapeutic genes into specific chromosomal sites using engineered retroviral integrase 使用工程逆转录病毒整合酶将治疗性基因安全递送到特定的染色体位点

Gene therapy and regulation Pub Date : 2004-12-01 DOI: 10.1163/1568558043967490

T. A. Wilkinson, W. Tan, S. A. Chow

{"title":"Safe delivery of therapeutic genes into specific chromosomal sites using engineered retroviral integrase","authors":"T. A. Wilkinson, W. Tan, S. A. Chow","doi":"10.1163/1568558043967490","DOIUrl":"https://doi.org/10.1163/1568558043967490","url":null,"abstract":"Gene therapy approaches that involve the permanent insertion of therapeutic genes into host chromosomal DNA have many desirable features and show considerable promise for success in the clinic. One major drawback of these approaches is that any unintended insertion events from the therapy can potentially have detrimental effects in patients, as demonstrated by the development of malignancies in both animal and human studies. Therefore, directing the integration of foreign genes into \"safe sites\" within the genome is highly desirable for these approaches. In retroviral-based vector systems, the viral enzyme integrase (IN) catalyzes the insertion of a desired transgene nonspecifically into the host cell genome. Efforts to engineer IN to recognize specific target DNA sequences within the genome, and thereby improve the safety of future generations of retroviral-based vectors, are described. Recent results using fusion protein constructs of IN and E2C, a designed polydactyl zinc-finger protein that specifically recognizes an 18-base pair DNA sequence, are highlighted in this review. Encouraging results have been generated in vitro, and additional studies are ongoing in mammalian cell systems. The long-term goal of these efforts is the development of effective retroviral vectors that can safely deliver therapeutics in a gene therapy setting.","PeriodicalId":93646,"journal":{"name":"Gene therapy and regulation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1163/1568558043967490","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"64795991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Editorial — Zinc finger nuclease-boosted gene targeting: toward clinical gene repair/alteration and custom site-specific integrative gene therapy 编辑-锌指核酸酶促进基因靶向:临床基因修复/改变和定制位点特异性综合基因治疗

Gene therapy and regulation Pub Date : 2004-08-01 DOI: 10.1163/1568558042457497

R. Bertolotti

引用次数: 2