新型基因表达调控元件——人工核糖开关的构建及筛选

核糖开关作为一种新发现的RNA元件,可以高效、准确、快速地执行基因调控任务,且免疫原性低,有可能在将来以顺式模块的方式应用于未来的基因治疗。近年来已经成功构建了多种人造核糖开关,构建方法主要是利用人工适体元件与基因表达调控元件组装,或者是在天然核糖开关基础上进行改造。文中全面综述了涉及人工核糖开关设计及筛选的技术,讨论了可以用于哺乳细胞、响应非天然配体信号、调控特征为热力学和动力学控制的核糖开关的设计新策略,并对核糖开关的筛选构建策略及其在基因治疗及新型药物开发领域的应用前景进行了展望。尽管目前将核糖开关设计成为功能强大的新型基因调控系统还面临很大的困难,但通过构效关系的研究、计算机辅助设计、体外筛选及细胞内筛选技术、高通量优化筛选等技术的综合应用,核糖开关一定可以成为有力的基因调控工具,如能成功应用则可大大促进基因治疗临床化的进程。     

RNA structure: Merging chemistry and genomics for a holistic perspective

The advent of deep sequencing technology has unexpectedly advanced our structural understanding of molecules composed of nucleic acids. A significant amount of progress has been made recently extrapolating the chemical methods to probe RNA structure into sequencing methods. Herein we review some of the canonical methods to analyze RNA structure, and then we outline how these have been used to probe the structure of many RNAs in parallel. The key is the transformation of structural biology problems into sequencing problems, whereby sequencing power can be interpreted to understand nucleic acid proximity, nucleic acid conformation, or nucleic acid‐protein interactions. Utilizing such technologies in this way has the promise to provide novel structural insights into the mechanisms that control normal cellular physiology and provide insight into how structure could be perturbed in disease.     

Computational Approaches for RNA Structure Ensemble Deconvolution from Structure Probing Data

RNA structure probing experiments have emerged over the last decade as a straightforward way to determine the structure of RNA molecules in a number of different contexts. Although powerful, the ability of RNA to dynamically interconvert between, and to simultaneously populate, alternative structural configurations, poses a nontrivial challenge to the interpretation of data derived from these experiments. Recent efforts aimed at developing computational methods for the reconstruction of coexisting alternative RNA conformations from structure probing data are paving the way to the study of RNA structure ensembles, even in the context of living cells. In this review, we critically discuss these methods, their limitations and possible future improvements.     

Engineering genes for predictable protein expression

The DNA sequence used to encode a polypeptide can have dramatic effects on its expression. Lack of readily available tools has until recently inhibited meaningful experimental investigation of this phenomenon. Advances in synthetic biology and the application of modern engineering approaches now provide the tools for systematic analysis of the sequence variables affecting heterologous expression of recombinant proteins. We here discuss how these new tools are being applied and how they circumvent the constraints of previous approaches, highlighting some of the surprising and promising results emerging from the developing field of gene engineering.     

反义RNA技术在丝状真菌代谢工程中的应用

丝状真菌作为一种重要的工业微生物,采用各种表达调控技术对其代谢途径进行改造以便适应生产需求成为当前的研究热点之一。反义RNA技术是代谢工程中调控基因表达的一种重要手段,且由于其操作简单避免了基因敲除技术的复杂性,在丝状真菌体系中有着良好的应用前景。本综述中,从反义RNA的作用机理、真菌体系的基因工程技术以及目前反义RNA技术的应用等方面,对反义RNA技术在丝状真菌代谢工程中的应用进行了概述。     

植物RNA病毒载体构建策略

作为对传统的植物转化载体的补充,植物ＲＮＡ病毒载体具有受体植物广泛,且转化需要的时间短,特别适宜于大量表达外源基因．虽然外源基因的稳定性存在一定的问题,但大量的事例说明,作为一种新型的外源基因在植物中表达的载体,它具有其独到的特点．本文就最新利用ＲＮＡ病毒转化植物的报道进行了综述