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Recent patents in RNA silencing in plants: constructs, methods and applications in plant biotechnology. 植物RNA沉默新专利:构建、方法及在植物生物技术中的应用。
Pub Date : 2010-11-01 DOI: 10.2174/187221510794751677
Sara López-Gomollón, Tamas Dalmay

RNA silencing is a recently discovered mechanism to regulate gene expression at transcriptional and posttranscriptional levels. It is based on the recognition and methylation of target genes or cleavage of target mRNAs by small RNA molecules, with length varying from 21 to 24 nucleotides. RNA silencing plays an important role modulating most of the important cell processes, such as growth, development or stress response. During the past few years, diverse strategies have been applied to exploit RNA silencing as a tool to create plants with enhanced economical properties or able to cope with pathogens or abiotic stress. This review describes the most important patents related to RNA silencing in plants, which disclose vectors designed to induce RNA silencing by hairpin RNAs, amplicons or virus-based plasmids, methods for detection and quantification of silencing as well as general uses in plant biotechnology.

RNA沉默是最近发现的在转录和转录后水平调节基因表达的机制。它是基于小RNA分子对靶基因的识别和甲基化或靶mrna的切割,长度从21到24个核苷酸不等。RNA沉默在调节大多数重要的细胞过程(如生长、发育或应激反应)中起着重要作用。在过去的几年中,各种策略被应用于利用RNA沉默作为一种工具来创造具有增强经济特性或能够应对病原体或非生物胁迫的植物。本文综述了植物中与RNA沉默相关的最重要专利,包括通过发夹RNA、扩增子或基于病毒的质粒诱导RNA沉默的载体、沉默的检测和定量方法以及在植物生物技术中的一般应用。
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引用次数: 5
RNA silencing. RNA silencing。
Pub Date : 2010-11-01 DOI: 10.2174/187221510794751631
Francisco E Nicolas
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引用次数: 0
Revealing biological information using data structuring and automated learning. 利用数据结构和自动学习揭示生物信息。
Pub Date : 2010-11-01 DOI: 10.2174/187221510794751668
Irina Mohorianu, Vincent Moulton

The intermediary steps between a biological hypothesis, concretized in the input data, and meaningful results, validated using biological experiments, commonly employ bioinformatics tools. Starting with storage of the data and ending with a statistical analysis of the significance of the results, every step in a bioinformatics analysis has been intensively studied and the resulting methods and models patented. This review summarizes the bioinformatics patents that have been developed mainly for the study of genes, and points out the universal applicability of bioinformatics methods to other related studies such as RNA interference. More specifically, we overview the steps undertaken in the majority of bioinformatics analyses, highlighting, for each, various approaches that have been developed to reveal details from different perspectives. First we consider data warehousing, the first task that has to be performed efficiently, optimizing the structure of the database, in order to facilitate both the subsequent steps and the retrieval of information. Next, we review data mining, which occupies the central part of most bioinformatics analyses, presenting patents concerning differential expression, unsupervised and supervised learning. Last, we discuss how networks of interactions of genes or other players in the cell may be created, which help draw biological conclusions and have been described in several patents.

在输入数据中具体化的生物学假设和通过生物学实验验证的有意义的结果之间的中间步骤通常使用生物信息学工具。从数据存储开始,到结果意义的统计分析结束,生物信息学分析的每一步都得到了深入研究,所产生的方法和模型获得了专利。本文综述了主要用于基因研究的生物信息学专利,并指出生物信息学方法在RNA干扰等相关研究中的普遍适用性。更具体地说,我们概述了在大多数生物信息学分析中所采取的步骤,重点介绍了从不同角度揭示细节的各种方法。首先,我们考虑数据仓库,这是必须有效执行的第一个任务,优化数据库的结构,以便于后续步骤和信息检索。接下来,我们回顾数据挖掘,它占据了大多数生物信息学分析的中心部分,提出了关于差异表达、无监督学习和有监督学习的专利。最后,我们讨论了如何在细胞中创建基因或其他参与者的相互作用网络,这有助于得出生物学结论,并已在几项专利中进行了描述。
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引用次数: 1
Recent patents and challenges on DNA microarray probe design technologies. DNA微阵列探针设计技术的最新专利与挑战。
Pub Date : 2010-11-01 DOI: 10.2174/187221510794751640
Dan Tulpan

The invention of microarray technology has empowered scientists to quickly transition from single gene studies to massively parallel experiments investigating thousands of genes. The use of DNA microarrays relies on accurate design for probes that are immobilized on a surface and bind specifically to complementary targets in a complex solution. The quality of a set of DNA probes heavily relies on DNA hybridization - the process of joining two single-strands of DNA to form a double-stranded molecule, and is traditionally ensured by using specific design criteria. The design of DNA probes for microarrays requires very stringent criteria, due to the necessity of choosing unique sequences that perfectly complement specific regions from large genomic data sets, while avoiding hybridization with every other region of the same genome. Patents and research publications presenting various probe design methods are reviewed in this manuscript and future potential extensions of current technologies are suggested.

微阵列技术的发明使科学家能够迅速从单基因研究过渡到大规模并行实验,研究数千个基因。DNA微阵列的使用依赖于探针的精确设计,这些探针固定在表面上,并在复杂溶液中特异性地与互补目标结合。一组DNA探针的质量很大程度上依赖于DNA杂交——将两条单链DNA连接起来形成双链分子的过程,传统上是通过使用特定的设计标准来保证的。微阵列DNA探针的设计需要非常严格的标准,因为必须选择独特的序列,以完美地补充大型基因组数据集中的特定区域,同时避免与同一基因组的所有其他区域杂交。专利和研究出版物提出了各种探针设计方法,在这篇论文中进行了审查,并提出了当前技术的未来潜在扩展。
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引用次数: 5
Recent patents of nanopore DNA sequencing technology: progress and challenges. 纳米孔DNA测序技术的最新专利:进展与挑战。
Pub Date : 2010-11-01 DOI: 10.2174/187221510794751622
Jianfeng Zhou, Bingqian Xu

DNA sequencing techniques witnessed fast development in the last decades, primarily driven by the Human Genome Project. Among the proposed new techniques, Nanopore was considered as a suitable candidate for the single DNA sequencing with ultrahigh speed and very low cost. Several fabrication and modification techniques have been developed to produce robust and well-defined nanopore devices. Many efforts have also been done to apply nanopore to analyze the properties of DNA molecules. By comparing with traditional sequencing techniques, nanopore has demonstrated its distinctive superiorities in main practical issues, such as sample preparation, sequencing speed, cost-effective and read-length. Although challenges still remain, recent researches in improving the capabilities of nanopore have shed a light to achieve its ultimate goal: Sequence individual DNA strand at single nucleotide level. This patent review briefly highlights recent developments and technological achievements for DNA analysis and sequencing at single molecule level, focusing on nanopore based methods.

DNA测序技术在过去几十年见证了快速发展,主要是由人类基因组计划推动的。在提出的新技术中,纳米孔被认为是超高速和极低成本的单DNA测序的合适候选技术。已经开发了几种制造和修饰技术来生产坚固且定义良好的纳米孔器件。在应用纳米孔分析DNA分子性质方面也做了许多努力。通过与传统测序技术的比较,纳米孔在样品制备、测序速度、成本效益和读取长度等主要实际问题上显示出了明显的优势。尽管挑战依然存在,但近年来在提高纳米孔能力方面的研究已经为实现其最终目标——在单核苷酸水平上对单个DNA链进行测序提供了线索。本专利综述简要介绍了单分子水平DNA分析和测序的最新进展和技术成就,重点是基于纳米孔的方法。
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引用次数: 5
Recent patents of DNA methylation biomarkers in gastrointestinal oncology. 胃肠道肿瘤DNA甲基化生物标志物新专利。
Pub Date : 2010-11-01 DOI: 10.2174/187221510794751695
Alejandro H Corvalan, Maria J Maturana

Gastrointestinal malignancies are among the most common malignancies worldwide. Advances in technology and treatment have improved diagnosis and monitoring of these tumors. As a consequence, identification of new biomarkers that can be applied at different levels of disease is urgently needed. DNA methylation is a process in which cytosines acquire a methyl group in 5' position only if they are followed by a guanine. An emerging catalog of specific genes inactivated by DNA methylation in gastrointestinal tumors has been established. In this review we will give a brief overview of the main sources of DNA used to investigate methylation biomarkers and several related patents. One of these is related to multiple genes that predict the risk of development of esophageal adenocarcinoma. Another evaluated methylation status of 24 genes to find one frequently methylated in primary tumors as well as plasma samples from gastric cancer patients. Others patented the epigenetic silencing of miR-342 as a promissory biomarker for colorectal carcinoma. Thus the new field of DNA methylation biomarkers holds the promise of better methods for screening, early detection, disease progression and outcome predictor of therapy response in gastrointestinal oncology.

胃肠道恶性肿瘤是世界上最常见的恶性肿瘤之一。技术和治疗的进步改善了这些肿瘤的诊断和监测。因此,迫切需要鉴定可以应用于不同疾病水平的新生物标志物。DNA甲基化是一个过程,在这个过程中,只有当胞嘧啶后面有一个鸟嘌呤时,胞嘧啶才能在5'位置上获得一个甲基。已经建立了胃肠道肿瘤中DNA甲基化失活的特定基因的新目录。在这篇综述中,我们将简要概述用于甲基化生物标志物研究的DNA的主要来源和几个相关专利。其中之一与预测食管癌发展风险的多个基因有关。另一项研究评估了24个基因的甲基化状态,发现一个基因在原发性肿瘤和胃癌患者的血浆样本中经常甲基化。其他人则申请了miR-342的表观遗传沉默作为结直肠癌的预期生物标志物的专利。因此,DNA甲基化生物标志物的新领域有望为胃肠道肿瘤的筛查、早期检测、疾病进展和治疗反应的结果预测提供更好的方法。
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引用次数: 15
MicroRNAs in human diseases. 人类疾病中的microrna。
Pub Date : 2010-11-01 DOI: 10.2174/187221510794751659
Francisco E Nicolas, Alfonso F Lopez-Martinez

The discovery of microRNAs (miRNAs), a new class of negative regulator that represses gene expression by pairing with their target messenger RNAs (mRNAs), has revealed a natural pathway for controlling gene expression. There are hundreds of miRNAs encoded in the human genome and thousands of target mRNAs, which illustrates the important regulatory roles of miRNAs in cell developmental, differentiation, proliferation and apoptosis pathways. In this scenario, it is not surprising that deregulated miRNAs have been involved in the pathogenesis of many human diseases. The recent development of technologies and compounds to identify and modulate miRNAs has opened new avenues for diagnosis, prognosis and therapeutic applications. Here, we summarize most of the recent patents related to the detection and profiling of miRNAs from pathological samples and to miRNA modulators used as new therapies for disease, including cancer and viral infections, as well as methods for their delivery.

microRNAs (miRNAs)是一类新的负调节因子,通过与其靶信使rna (mrna)配对来抑制基因表达,它的发现揭示了控制基因表达的自然途径。人类基因组编码的mirna有数百种,靶mrna有数千种,这说明了mirna在细胞发育、分化、增殖和凋亡等途径中的重要调控作用。在这种情况下,不受调控的mirna参与许多人类疾病的发病机制就不足为奇了。近年来,鉴定和调节mirna的技术和化合物的发展为诊断、预后和治疗应用开辟了新的途径。在这里,我们总结了最近与病理样本中miRNA的检测和分析相关的大多数专利,以及用于疾病(包括癌症和病毒感染)新疗法的miRNA调节剂,以及它们的递送方法。
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引用次数: 29
Patented applications of gene silencing in plants: manipulation of traits and phytopathogen resistance. 基因沉默在植物中的专利应用:性状操纵和植物病原菌抗性。
Pub Date : 2010-11-01 DOI: 10.2174/187221510794751613
Ruben Alvarez-Fernandez

RNA silencing is the name of a broad family of phenomena including RNA interference (RNAi) in animals and basal eukaryotes, quelling in fungi and posttranscriptional gene silencing (PTGS) in plants. PTGS is a fertile research field and since its discovery many applications have been developed related to plant breeding. This minireview summarizes those patents which apply engineered gene silencing to specific problems. The range of inventions is divided in two main sections: manipulation of traits and resistance to phytopathogens and pests. Subtopics like manipulation of tolerances to abiotic stress, alteration of lignin, biofactories, alkaloids biosynthesis and flowering time fall within the first section, and introduction of resistances to insects, nematodes, bacteria, virus and fungi can be found within the second one.

RNA沉默是一系列广泛现象的统称,包括动物和基础真核生物中的RNA干扰(RNAi)、真菌中的抑制(quelling)和植物中的转录后基因沉默(PTGS)。PTGS是一个丰富的研究领域,自从它被发现以来,已经开发了许多与植物育种有关的应用。本文综述了应用工程基因沉默技术解决具体问题的专利。发明范围分为两个主要部分:性状操纵和对植物病原体和害虫的抗性。诸如操纵对非生物胁迫的耐受性、木质素的改变、生物工厂、生物碱的生物合成和开花时间等子主题属于第一部分,而对昆虫、线虫、细菌、病毒和真菌的抗性的引入可以在第二部分找到。
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引用次数: 1
Strategies for detecting genomic DNA methylation: a survey of US patents. 检测基因组DNA甲基化的策略:美国专利调查。
Pub Date : 2010-06-01 DOI: 10.2174/187221510793205719
M Ahmad Chaudhry

DNA methylation is one of the mechanisms for the epigenetic control of gene expression. Alterations in the methylation status of genomic DNA can result in the silencing of genes. Such control is of significance for a wide range of biological processes, ranging from cellular differentiation during development, genomic imprinting and X-chromosome inactivation to the maintenance of genome stability. The cytosine in the genomic DNA is converted to 5-methylcytosine. The hypermethylation of some CpG islands in genomic DNA could result in gene silencing and hypomethylation can lead to transcription and gene expression. There has been a great interest in developing molecular techniques to analyze genomic DNA methylation at the CpG islands. The discovery that DNA treatment with sodium bisulfite converts the cytosine to uracil while keeping the 5-methycytosine intact has opened the door to a number of strategies to investigate genomic DNA methylation both at regional and global levels. A survey of recently patented methods to analyze DNA methylation indicated a range of inventions from simple PCR to high throughput based technologies. The disease diagnosis was the prominent application of DNA methylation detection for most of these methods. Future inventions will likely concentrate on genome-scale DNA methylation discovery.

DNA甲基化是基因表达的表观遗传调控机制之一。基因组DNA甲基化状态的改变可导致基因沉默。这种控制对广泛的生物过程具有重要意义,从发育过程中的细胞分化,基因组印迹和x染色体失活到基因组稳定性的维持。基因组DNA中的胞嘧啶转化为5-甲基胞嘧啶。基因组DNA中某些CpG岛的高甲基化可导致基因沉默,而低甲基化可导致转录和基因表达。人们对开发分子技术来分析CpG岛的基因组DNA甲基化非常感兴趣。亚硫酸氢钠处理DNA将胞嘧啶转化为尿嘧啶,同时保持5-甲基胞嘧啶的完整,这一发现为在区域和全球水平上研究基因组DNA甲基化的许多策略打开了大门。一项对最近获得专利的DNA甲基化分析方法的调查显示,从简单的PCR到高通量技术的一系列发明。在这些方法中,DNA甲基化检测在疾病诊断中的应用最为突出。未来的发明可能会集中在基因组尺度的DNA甲基化发现上。
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引用次数: 3
Recent patents on immunoregulatory DNA vaccines for autoimmune diseases and allograft rejection. 自身免疫疾病和同种异体移植排斥免疫调节DNA疫苗的最新专利。
Pub Date : 2010-06-01 DOI: 10.2174/187221510793205700
Shahrokh Shabahang, Alice F Li, Alan Escher

The goal of immunoregulatory DNA vaccination is the antigen- and tissue-specific suppression of pathological inflammation that underlies immune-mediated inflammatory disorders like autoimmune diseases and allograft rejection. Recent patents and patent applications have applied immunoregulatory DNA vaccines in rodent model systems and human clinical trials using plasmid DNA coding for autoantigens such as insulin and glutamic acid decarboxylase for type 1 diabetes, myelin-associated proteins for multiple sclerosis, and heat-sock protein 60 for rheumatoid arthritis. In these cases, the objective is to induce a homeostatic-like regulatory immune response to suppress pathological inflammation. In addition, patent applications have disclosed the use of DNA vaccines encoding the pro-inflammatory MIF cytokine and the CD25 IL-2 receptor subunit to interfere with the inflammatory process. Approaches have also been taken to improve DNA vaccination efficacy, including covalent modification of plasmid DNA, engineering secretion of vaccine-encoded antigen, and co-delivery of DNA coding for anti-inflammatory cytokines, a mutant co-stimulatory molecule, a growth factor, or a pro-apoptotic protein. Furthermore, a patent application has disclosed the use of a DNA vaccine previously shown to treat successfully an autoimmune disease to prolong allograft survival. Taken together, these patents and patent applications indicate a promising bench-to-bedside potential for immunoregulatory DNA vaccination applied to autoimmune diseases and allograft rejection.

免疫调节性DNA疫苗的目标是抗原和组织特异性抑制病理性炎症,这是免疫介导的炎症疾病如自身免疫性疾病和同种异体移植排斥反应的基础。最近的专利和专利申请已将免疫调节DNA疫苗应用于啮齿动物模型系统和人类临床试验中,使用质粒DNA编码自身抗原,如用于1型糖尿病的胰岛素和谷氨酸脱羧酶,用于多发性硬化症的髓磷脂相关蛋白和用于类风湿性关节炎的热sock蛋白60。在这些情况下,目的是诱导一种类似于体内平衡的调节免疫反应来抑制病理性炎症。此外,专利申请已经披露了使用编码促炎MIF细胞因子和CD25 IL-2受体亚基的DNA疫苗来干扰炎症过程。提高DNA疫苗接种效果的方法也已被采用,包括质粒DNA的共价修饰,疫苗编码抗原的工程分泌,以及抗炎细胞因子、突变共刺激分子、生长因子或促凋亡蛋白编码DNA的共同递送。此外,一项专利申请披露了先前显示可成功治疗自身免疫性疾病以延长同种异体移植物存活的DNA疫苗的使用。综上所述,这些专利和专利申请表明,将免疫调节DNA疫苗应用于自身免疫性疾病和同种异体移植排斥反应具有从实验到临床的巨大潜力。
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引用次数: 3
期刊
Recent patents on DNA & gene sequences
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