Mechanistic studies for the role of cellular nucleic-acid-binding protein (CNBP) in regulation of c-myc transcription
Siqi ChenLijuan SuJun QiuNannan XiaoJing LinJia‐Heng TanTian‐Miao OuLian‐Quan GuZhi‐Shu HuangDing Li
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Gene silencing is one of the important factors of transgene inactivation. The main mechanisms, including postion effects, transcriptional silencing and post-transcriptional silencing was presented, the means of stabilizing gene expression in transgeneic plants was discussed.
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This chapter contains sections titled: Introduction Gene Silencing—The Basic Parts List Mechanisms of Silencing Gene Silencing in Oomycetes: Tales from the Laboratory and Clues from Genomes Strategies for Application of Gene Silencing in Oomycetes Validation: Linking Gene Silencing to Phenotype Which Strategy? Stable versus Transient Gene Silencing Conclusions and Scope for Future Work Acknowledgments References
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Gene silencing is one of the important factors of transgene inactivation. The main mechanisms , including postion effects, transcriptional silencing and post transcriptional silencing was presented , the means of stabilizing gene expression in transgeneic plants was discussed.
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Intracellular siRNA release is a crucial step in efficient gene silencing mediated by cationic polymers. Here, we show an example of temperature change-induced intracellular siRNA release and silencing using a temperature-responsive polymer consisting of dendrimer, poly(N-isopropylacrylamide) and phenylboronic acid. The smart polymer can trigger the release of loaded siRNA in a controlled manner upon cooling the surrounding solution below its lower critical solution temperature. Gene silencing efficacy of the polymer was significantly increased by cool treatment after its cellular uptake. The polymer and the cool treatment cause minimal toxicity to the transfected cells. The results provide a facile and promising strategy to design stimuli-responsive polymers for efficient gene silencing.
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We showed previously that grafting transmitted silencing occurred when transgenic ACC oxidase 1 (ACO1) overexpressing tomato plants that also produced siRNAs were grafted onto transgenic stocks that already showed strong silencing. The presence of siRNAs in these overexpressing scions may indicate that silencing, though inefficient, may already occur at a low level before grafting. To test if a silencing state with a relatively high level of target mRNA can be shifted towards further more effective silencing, we grafted an ACO1 antisense (AS) line with a high level of antisense ACO1 transgene mRNA and low level of siRNAs to the ACO1 strong silencer stock. The AS mRNA level was reduced dramatically two weeks after grafting. More interestingly, self-grafting of ACO1 overexpressers and AS lines also induced strong silencing in the scions. We suggest that grafting transmitted silencing may involve the switching from an inefficient or weak silencing state to a stronger silencing by a systemic silencing signal, similar to the change of silencing states that sometimes occurs during development. Control experiments using non-transgenic stocks designed to test whether wounding alone is responsible for generating a signal that enhances silencing in transgenic scions gave negative results. We propose that the build-up of silencing signal and / or molecules at both sides of the grafting junction and their sudden release when the phloem is reconnected may be critical to grafting transmitted silencing.
RNA Silencing
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Transgenes are inactive and the trans-gene expression couldn't be detected thoroughly ,which are called as gene silencing in transgenet-ic plants. The main mechanisms, including position effects, transcriptional silencing and post-transcriptional silencing was presented, the means of stabilizing gene expression in transge-netic plants was discussed. Studying the cause for silencing of foreign genes and learning the method to overcome gene silencing are important for the development of plant gene engineering.
Gene engineering
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