细胞友好的新型基因导入法

基因导入，是现代生命科学研究中的基础性技术。正因为如此，以病毒载体法为首，粒递送法以及电穿孔法等已经确立的有很多方法。BIOBANK（日本东京都港区）公司正在销售一种新型基因导入装置“共振激发式基因导入装置”，与原有的那些方法相比，它能够高效地导入基因。公司正在大力宣传其优点，努力打入基础研究的过程中。     

叶绿体遗传转化：植物导入外源基因的新途径

叶绿体转化系统,独立于传统的核转化,为植物导入外源基因提供了新途径,它有如下优点：超量表达目的的基因;以定点整合方式导入外源基因从而消除了位置效应及基因沉默;具有原核表达方式,能以多 反子的形式表达多个基因;母系遗传方式可防止基因扩散;基因产物区域化并能提供适于某些产物发挥功能的小环境等。随着这项技术在越来越多的领域发挥作用,其优势性逐渐得到认同。本文着重对此技术的特点,发展及应用作一综述。     

外源基因的靶向细胞表达

使外源基因在特定组织细胞中进行表达主要采用两方：一是使用靶向基因载体，这种载体可识别特定组织细胞膜上的一些分子，并与之结合，然后再将外源基因带入细胞中表达；另一种方式是使用组织细胞专一的启动子，来驱动外源基因在该细胞中的特异性表达。     

将外源基因导入水稻的研究现状及其展望

水稻是世界上最重要的农作物之一,全球有近一半的人口以水稻作为主粮。水稻育种一直是各国育种学家最重视的研究课题。目前,水稻育种实践中仍主要采用杂交育种等常规育种手段。但是近十年来,随着生物技术的迅速发展,各国育种学家愈来愈广泛地采用基因工程等现代生物技术于水稻育种研究中,试图将一些控制优良性状的外源基因导入水稻,从而培育出高产、优质、抗逆性强的水稻新品种。这一研究领域最近几年取得了突破性进展。 (一)将外源基因导入水稻的途径 近十年来,植物基因工程取得了突飞猛进的发展。据统计,迄今已获得40余种植物的转基因植物,其中包括黄瓜、烟草、棉花、大豆、马铃薯、番茄及向日葵等十余种重要经济作物。上述植物的转基因植物的获得,在很大程度上是依赖基因工程载体——根癌农杆Ti菌质粒载体的。但是,水稻基因工程育种则一开始就遇上了难题,即现有的根癌农杆菌Ti质粒载体不适用于水稻,原因是根癌农杆菌不能感染作为禾本科植物之一的     

基因治疗导入载体的研究进展

基因治疗在恶性肿瘤、癌症、遗传性疾病和心脑血管等疾病的治疗中开始应用,临床治疗效果明显。基因治疗中的关键技术是选用合适的载体将外源基因高效导入受体靶细胞,综述了基因治疗中病毒和非病毒载体的研究进展。     

活体电穿孔法基因导入技术

活体电穿孔法(invivoelectroporation)可将外源基因有效导入靶组织或器官,导入效率较高,并且可在多种组织器官上应用。近年来活体电穿孔法用于转基因研究的报道不断增多,在基因治疗方面的优势也日趋显著,是一种很好的活体基因导入方法 。     

基因治疗的现状与展望

自从明确了单基因缺陷的疾病,就提出了基因治疗的概念。基因治疗是直接通过基因传递治疗人类疾病的一种手段,还可以理解成临床症状通过基因物质的传递而行到改善。目前在这个领域已经投入了几十亿美元,开展了超过三百例的临床试验,人们正通过不懈努力,期待着基因治疗的美好前景。     

病毒--基因治疗中有效的载体系统

基因治疗面临的首要问题是如何选择适当的基因载体将具有治疗价值的基因导入靶细胞并使其有效表达,以达到治疗疾病的目的。目前基因治疗临床试验中采用的载体大多数为病毒载体。本文主要介绍基因治疗中常用的4种病毒载体的生物学特性,以及各个载体在基因治疗中的优缺点。     

高赖氨酸蛋白基因导入水稻及可育转基因植株的获得

构建了一个植物高效表达质粒,使来源于四棱豆（Psophocarpus tetragonolobus(L.)DC)的高赖氨酸蛋白基因（lys）受控于单子叶植物ubiqutin强启动子下表达。用基因枪法将其导入水稻(Oryza sativa L.)幼胚诱导的愈伤组织,经潮霉素抗性筛选,得到可育的再生植株。经PCR和Southem blotting检测,表明该基因已整合到水稻的基因组织。GUS组织化学染色表明转基因水稻植株的叶、茎和根中均有gus基因的表达。测定112株转基因水稻叶片中赖氨酸叶量,大部分植株有不同程度的提高,最高幅度为16．04％。     

外源DNA(基因)导入大豆的研究

大豆(Glycine max),豆科,各地均有栽培[1].世界各国许多科技工作者都致力于大豆新品种的选育和大豆品质改良方面的研究工作,尤其致力于先进育种技术和手段的研究与应用.常规育种技术是有性杂交、回交和轮回选择等,有极大局限性[2].     

Gene delivery methods in cardiac gene therapy

Gene therapy for the treatment of heart failure is emerging as a multidisciplinary field demonstrating advances with respect to identifying key signaling pathways, modernized vector creation and delivery technologies. Although these discoveries offer significant progress, selecting optimal methods for the vector delivery remains a key component for efficient cardiac gene therapy to validate the targets in rodent models and to test clinically relevant ones in pre-clinical models. Although the goals of higher transduction efficiency and cardiac specificity can be achieved with several delivery methods, the invasiveness and patient safety remain unclear for clinical application. In this review, we discuss various features of the currently available vector delivery methods for cardiac gene therapy.     

Cationic liposome-mediated gene delivery in vivo

Several improvements have been made in liposomal delivery, thus making this technology potentially useful for treatment of certain diseases in the clinic. Success in non-viral delivery is complicated and requires optimization of several components. These components include nucleic acid purification, plasmid design, formulation of the delivery vehicle, administration route and schedule, dosing, detection of gene expression, and others. With further improvements, broad use of non-viral delivery systems to treat human disorders should be possible.     

Engineering protein self-assembling in protein-based nanomedicines for drug delivery and gene therapy

Lack of targeting and improper biodistribution are major flaws in current drug-based therapies that prevent reaching high local concentrations of the therapeutic agent. Such weaknesses impose the administration of high drug doses, resulting in undesired side effects, limited efficacy and enhanced production costs. Currently, missing nanosized containers, functionalized for specific cell targeting will be then highly convenient for the controlled delivery of both conventional and innovative drugs. In an attempt to fill this gap, health-focused nanotechnologies have put under screening a growing spectrum of materials as potential components of nanocages, whose properties can be tuned during fabrication. However, most of these materials pose severe biocompatibility concerns. We review in this study how proteins, the most versatile functional macromolecules, can be conveniently exploited and adapted by conventional genetic engineering as efficient building blocks of fully compatible nanoparticles for drug delivery and how selected biological activities can be recruited to mimic viral behavior during infection. Although engineering of protein self-assembling is still excluded from fully rational approaches, the exploitation of protein nano-assemblies occurring in nature and the direct manipulation of protein–protein contacts in bioinspired constructs open intriguing possibilities for further development. These methodologies empower the construction of new and potent vehicles that offer promise as true artificial viruses for efficient and safe nanomedical applications.     

The evolution of heart gene delivery vectors

Gene therapy holds promise for treating numerous heart diseases. A key premise for the success of cardiac gene therapy is the development of powerful gene transfer vehicles that can achieve highly efficient and persistent gene transfer specifically in the heart. Other features of an ideal vector include negligible toxicity, minimal immunogenicity and easy manufacturing. Rapid progress in the fields of molecular biology and virology has offered great opportunities to engineer various genetic materials for heart gene delivery. Several nonviral vectors (e.g. naked plasmids, plasmid lipid/polymer complexes and oligonucleotides) have been tested. Commonly used viral vectors include lentivirus, adenovirus and adeno-associated virus. Among these, adeno-associated virus has shown many attractive features for pre-clinical experimentation in animal models of heart diseases. We review the history and evolution of these vectors for heart gene transfer.     

基因兴奋剂

人类基因组计划的完成为基因治疗提供了依据,同时也给基因兴奋剂的产生创造了条件。本文通过综述基因兴奋剂的概念、产生的分子机制,讨论了可能被用作基因兴奋剂的基因,并从基因、转录和蛋白质三个水平探讨了基因兴奋剂的检测策略。     

Acid‐activated nonviral peptide vector for gene delivery

Nonviral vector–based gene therapy is a promising strategy for treating a myriad of diseases. Cell‐penetrating peptides are gaining increasing attention as vectors for nucleic acid delivery. However, most studies have focused more on the transfection efficiency of these vectors than on their specificity and toxicity. To obtain ideal vectors with high efficiency and safety, we constructed the vector stearyl‐TH by attaching a stearyl moiety to the N‐terminus of the acid‐activated cell penetrating peptide TH in this study. Under acidic conditions, stearyl‐TH could bind to and condense plasmids into nanoparticle complexes, which displayed significantly enhanced cellular uptake and transfection efficiencies. In contrast, stearyl‐TH lost the capacities of DNA binding and transfection at physiological pH. More importantly, stearyl‐TH and the complexes formed by stearyl‐TH and plasmids displayed no obvious toxicity at physiological pH. Consequently, the high transfection efficiency under acidic conditions and low toxicity make stearyl‐TH a potential nucleic acid delivery vector for gene therapy.     

Mesenchymal stem cells as carrier of the therapeutic agent in the gene therapy of blood disorders

Nonhematopoietic stem cells as a delivery platform of therapeutic useful genes have attracted widespread attention in recent years, owing to gained a long lifespan, easy separation, high proliferation, and high transfection capacity. Mesenchymal stem/stromal cells (MSCs) are the choice of the cells for gene and cell therapy due to high self-renewal capacity, high migration rate to the site of the tumor, and with immune suppressive and anti-inflammatory properties. Hence, it has a high potential of safety genetic modification of MSCs for antitumor gene expression and has paved the way for the clinical application of these cells to target the therapy of cancers and other diseases. The aim of gene therapy is targeted treatment of cancers and diseases through recovery, change, or enhancement cell performance to the sustained secretion of useful therapeutic proteins and induction expression of the functional gene in intended tissue. Recent developments in the vectors designing leading to the increase and durability of expression and improvement of the safety of the vectors that overcome a lot of problems, such as durability of expression and the host immune response. Nowadays, gene therapy approach is used by MSCs as a delivery vehicle in the preclinical and the clinical trials for the secretion of erythropoietin, recombinant antibodies, coagulation factors, cytokines, as well as angiogenic inhibitors in many blood disorders like anemia, hemophilia, and malignancies. In this study, we critically discuss the status of gene therapy by MSCs as a delivery vehicle for the treatment of blood disorders. Finally, the results of clinical trial studies are assessed, highlighting promising advantages of this emerging technology in the clinical setting.