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内参
内参在生物学研究中非常常见。例如RT-PCR或western blot实验中常用表达量稳定的管家基因/蛋白作为评估不同样本之间目标基因或蛋白变化的参考。以western blot为例，如果加药处理前后GAPDH或tublin、actin等基本没有变化，而目标蛋白表达变化明显，则说明“加药”过程中目标蛋白的表达受到影响。内参的引入可以排除上样量不同或样本处理过程中的偏差导致的可能误差···
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应用靶向种子序列的锚定核酸沉默整个miRNA家族
Loss-of-function是基因功能研究的常用方法之一。但是，在microRNA功能研究、特别是microRNA家族功能研究中，loss-of-function却遇到一些技术瓶颈。为此Obad S等建立了应用针对microRNA家族的8 mer种子序列的锚定核酸(locked ncleiuc acid，LNA)抑制所有包含该种子序列的microRNA 家族成员的新方法（Silencing of microRNA families by seed-targeting tiny LNAs Nature Genetics 2011，43，371-378）。在转染针对特定基因家族（例如miR21家族）种子序列的小LNA（tiny LNA）之后，发现相应的靶mRNA同时被抑制。这种非结合性的小LNA可以方便地导入各种正常组织或肿瘤组织（例如小鼠乳腺肿瘤）中，实现长时间的、针对特定miRNA家族的干涉效应。
图1 tiny LNA作用原理
Silencing of microRNA families by seed-targeting tiny LNAs
Abstract The challenge of understanding the widespread biological roles of animal microRNAs (miRNAs) has prompted the development of genetic and functional genomics technologies for miRNA loss-of-function studies. However, tools for exploring the functions of entire miRNA families are still limited. We developed a method that enables antagonism of miRNA function using seed-targeting 8-mer locked ncleiuc acid (LNA) oligonucleotides, termed tiny LNAs. Transfection of tiny LNAs into cells resulted in simultaneous inhibition of miRNAs within families sharing the same seed with concomitant upregulation of direct targets. In addition, systemically delivered, unconjugated tiny LNAs showed uptake in many normal tissues and in breast tumors in mice, coinciding with long-term miRNA silencing. Transcriptional and proteomic profiling suggested that tiny LNAs have negligible off-target effects, not significantly altering the output from mRNAs with perfect tiny LNA complementary sites. Considered together, these data support the utility of tiny LNAs in elucidating the functions of miRNA families in vivo.
原文链接：http://www.nature.com/ng/journal/v43/n4/full/ng.786.html
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