Nonviral transfection of mouse calvarial organ in vitro using Accell-modified siRNA.
Nonviral transfection of mouse calvarial organ in vitro using Accell-modified siRNA.
复制标题
使用 Accell 修饰的 siRNA 对小鼠颅骨器官进行体外非病毒转染。
DOI：
10.1097/prs.0b013e3181c82df1
复制
发表时间：
2010
影响因子：
3.6
通讯作者：
Gosain,ArunK
中科院分区：
文献类型：
--
作者：
Gupta,AshimK;Eshraghi,Yashar;Gliniak,Christy;Gosain,ArunK
背景：了解颅缝融合的生物学和参与该过程的分子的确切作用将有助于确定参与缝融合调节的关键因素。这些关键因素的调节可以作为一种组织工程技术，以取代传统的外科手术来纠正过早的缝线融合。通过RNA干扰调节基因表达是一种广泛使用的具有高潜力的技术。由于没有可用的报告，在体外的颅骨器官转染，作者研究了一个成功的非病毒递送技术的小抑制RNA（siRNA）在体外颅骨器官culturesystem.Methods：在这项研究中，19日龄的雄性CD 1小鼠被安乐死，并通过顶骨和额骨，2毫米的矢状缝两侧平行开颅手术，小心，以保持下面的硬脑膜的发展。结果：转化生长因子-β 1特异性Accell修饰的siRNA转染颅骨器官后可有效地降低mRNA水平。结论：本研究的观察结果表明，在体外颅骨器官培养系统中，使用Accell修饰的siRNA可获得特异、有效和持久的RNA干扰活性。除了绕过对有毒脂质载体的需要外，Accell修饰的siRNA中引入的修饰使其更稳定，更少脱靶。一旦确定了基因特异性siRNA对体外缝线融合的初始作用，这种技术就有可能用于体内研究。
Background:Understanding the biology of cranial suture fusion and the precise role of involved molecules implicated in the process will help to identify key factors involved in regulation of suture fusion. Modulation of these key factors may serve as a tissue-engineering technique to replace the traditional surgical procedures for the correction of premature suture fusion. Modulation of gene expression by RNA interference is a widely used technique with high potential. Because there is no available report of calvarial organ transfection in vitro, the authors studied the development of a successful nonviral delivery technique of small inhibitory RNA (siRNA) to an in vitro calvarial organ culture system.Methods:In this study, 19-day-old male CD1 mice were euthanized and parallel craniotomies made through the parietal and frontal calvaria, 2 mm to either side of the sagittal suture, with care taken to preserve the underlying dura mater. Organs grown in vitro in a defined medium were transfected with transforming growth factor-β1–specific Accell-modified siRNA followed by RNA isolation and quantitative polymerase chain reaction analysis.Results:Transfection of a calvarial organ with transforming growth factor-β1–specific Accell-modified siRNA effectively knocks down the mRNA level.Conclusions:Observations from this study indicate that in an in vitro calvarial organ culture system, a specific, efficient, and durable RNA interference activity can be achieved when Accell-modified siRNA is used. In addition to bypassing the need for toxic lipid carriers, the modifications introduced in Accell-modified siRNAs make it more stable and less off-target. This technique can potentially be used for in vivo studies once the initial effect of gene-specific siRNA on in vitro suture fusion has been determined.
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影响因子：
56.9
作者：
BERRY, SD;THOMPSON, RF
通讯作者：
THOMPSON, RF
经典条件反射的海马基质
DOI：
10.3758/bf03332858
发表时间：
1980
期刊：
Psychobiology
影响因子：
--
作者：
Richard F. Thompson;T. Berger;S. Berry;F. Hoehler;R. Kettner;D. Weisz
通讯作者：
D. Weisz
影响因子：
2.9
作者：
DISTERHOFT, JF;KWAN, HH;LO, WD
通讯作者：
LO, WD
海马神经元重复放电后会出现钙激活的超极化。
影响因子：
2.5
作者：
J. Hotson;D. Prince
通讯作者：
D. Prince
DOI：
10.3758/bf03326630
发表时间：
1979
期刊：
Psychobiology
影响因子：
--
作者：
JOHN W. Moore
通讯作者：
JOHN W. Moore