Correlations between recombination rate and intron distributions along chromosomes of C. elegans

Generally speaking, the intron size positively correlates with recombination rate in Caenorhabditis elegans genome. Here, we analyze the correlations between recombination rate and some measures of different intron lengths so as to know whether the recombination influences the introns of different lengths in the same way. Results show that the correlation between the recombination rate and the percentage of short introns (<100 bp) is negative, but the correlation between the recombination rate and the percentage of introns that are larger than 500 bp is positive. Average intron length correlates positively with the recombination rate for introns whose length is in the range of 100–1000 bp. We speculate that the recombination mainly exerts impact on introns whose length ranges from 100–1000 bp. We also show that the average intron number per gene correlates negatively with the recombination rate.     

基于核酶的自剪切sgRNA及其在基因编辑中的应用

为减少在CRISPR/Cas9基因编缉系统中,sgRNA通常由Ⅲ型启动子持续转录产生,Ⅲ型启动子不易控制,sgRNA持续表达又容易产生细胞毒性这一缺陷,本文拟建立由Ⅱ型启动子转录,并由内含子中释放sgRNA的方法.我们在sgRNA的两端设计了三种"核酶-sgRNA-核酶"的组合,通过核酶的自剪切作用实现sgRNA的释放.实验结果表明,HHRz-sgRNA-HDVRz的设计能够正确的从内含子中释放sgRNA,并且该sgRNA在构建的copGFP敲除模型中能够实现有效的基因编辑.这一发现证明在哺乳动物细胞中利用Ⅱ型启动子,在核酶和内含子剪切的协助下可以实现sgRNA的表达,这为Cas9和sgRNA整合在一个Ⅱ型启动子之下实现组织特异性和药物诱导性表达提供了基础.     

Cloning and evolutionary analysis of cyclin B gene introns in cyprinids with different ploidy levels

Partial fragments of the cyclin B gene from triploid, tetraploid, and pentaploid hybrids of red crucian carp × blunt snout bream, blunt snout bream, grass carp, silver carp, and bighead carp were amplified. One DNA fragment was amplified from the blunt snout bream, grass carp, silver carp, and bighead carp (750, 950, 720, and 720 bp, respectively). Two fragments (1200 and 900 bp) were amplified from the red crucian carp, common carp, and allotetraploids. The triploid and pentaploid hybrids yielded three DNA fragments (1200, 900, and 750 bp). The 1200 bp fragment of the allotetraploid crucian carp, triploid, tetraploid, pentaploid hybrids of red crucian carp × blunt snout bream shared 99.5%, 98.9%, 99.5%, and 88.7% homology, respectively, with the maternal DNA. The 900 bp fragment shared 97.5%, 94.6%, 94.2%, and 89.9% homology, respectively. Our results suggest that inheritance is maternally dominated. Furthermore, we observed preferential elimination of the paternal sequences in the allotetraploid hybrids. Based on these sequence analyses we constructed a phylogenetic tree to explain the relationships among the different ploidy levels.     

核仁小分子RNA及其基因组织形式

核仁小分子RNA(snoRNA)是非编码RNA中研究得最多,了解得最详细的成员之一。依据snoRNA保守的序列和结构元件可将其分成三类:box C/D snoRNA,box H/ACA snoRNA以及MRP RNA。它们的主要功能分别为指导rRNA或snRNA中特异位点的2′-O-核糖甲基化修饰、假尿嘧啶化修饰或作为分子伴侣参与靶标RNA高级结构的形成。目前发现的snoRNA基因组织主要有四种形式:独立基因编码的snoRNA、内含子编码的snoRNA、多顺反子snoRNA基因簇和内含子编码的snoRNA基因簇。但是,并不是每种生物都具有这四种基因组织形式。大多数脊椎动物的snoRNA产生于蛋白质基因的内含子中,植物snoRNA基因多以基因簇的形式存在,大部分酵母snoRNA由独立基因编码,锥体虫snoRNA以多顺反子形式存在,果蝇snoRNA基因都位于宿主基因的内含子中。随着对snoRNA研究工作的不断深入,snoRNA基因组织形式也呈现出前所未有的复杂性和多样性。