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The dynamic chromatin landscape and mechanisms of DNA methylation during mouse germ cell development. 小鼠生殖细胞发育过程中DNA甲基化的动态染色质景观和机制。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-04-16 DOI: 10.1266/ggs.21-00069
K. Shirane
Epigenetic marks including DNA methylation (DNAme) play a critical role in the transcriptional regulation of genes and retrotransposons. Defects in DNAme are detected in infertility, imprinting disorders and congenital diseases in humans, highlighting the broad importance of this epigenetic mark in both development and disease. While DNAme in terminally differentiated cells is stably propagated following cell division by the maintenance DNAme machinery, widespread erasure and subsequent de novo establishment of this epigenetic mark occur early in embryonic development as well as in germ cell development. Combined with deep sequencing, low-input methods that have been developed in the past several years have enabled high-resolution and genome-wide mapping of both DNAme and histone post-translational modifications (PTMs) in rare cell populations including developing germ cells. Epigenome studies using these novel methods reveal an unprecedented view of the dynamic chromatin landscape during germ cell development. Furthermore, integrative analysis of chromatin marks in normal germ cells and in those deficient in chromatin-modifying enzymes uncovers a critical interplay between histone PTMs and de novo DNAme in the germline. This review discusses work on mechanisms of the erasure and subsequent de novo DNAme in mouse germ cells as well as the outstanding questions relating to the regulation of the dynamic chromatin landscape in germ cells.
包括DNA甲基化(DNAme)在内的表观遗传标记在基因和反转录转座子的转录调控中起着至关重要的作用。在人类的不孕症、印记疾病和先天性疾病中检测到DNAme缺陷,突出了这种表观遗传标记在发育和疾病中的广泛重要性。虽然终分化细胞中的DNAme在细胞分裂后通过维持DNAme机制稳定地繁殖,但这种表观遗传标记的广泛清除和随后的重新建立发生在胚胎发育早期和生殖细胞发育中。结合深度测序,过去几年开发的低输入方法已经能够在包括发育中的生殖细胞在内的罕见细胞群体中实现DNAme和组蛋白翻译后修饰(PTMs)的高分辨率和全基因组定位。使用这些新方法的表观基因组研究揭示了生殖细胞发育过程中动态染色质景观的前所未有的观点。此外,对正常生殖细胞和缺乏染色质修饰酶的生殖细胞中染色质标记的综合分析揭示了组蛋白PTMs与生殖细胞中新生DNAme之间的关键相互作用。本文综述了在小鼠生殖细胞中DNAme的清除及其新生机制方面的研究工作,以及生殖细胞中染色质动态格局调控方面的悬而未决的问题。
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引用次数: 2
ANRIL as a prognostic biomarker in colon pre-cancerous lesion detection via non-invasive sampling. ANRIL作为无创取样检测结肠癌前病变的预后生物标志物。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-03-17 DOI: 10.1266/ggs.21-00102
Shadi Sadri, L. Rejali, M. Hadizadeh, H. A. Aghdaei, Chris Young, E. Nazemalhosseini-Mojarad, M. Zali, M. Bonab
Long non-coding RNAs have been proposed as biomarkers for the detection, prevention and screening of various malignancies. In this study, two lncRNAs (ANRIL and BANCR) were assessed for biomarker application in the early detection of colorectal cancer (CRC) through stool specimen testing, as a non-invasive and cost-effective methodology. A total of 40 stool samples were collected from patients referred to the hospital with colorectal cancer or adenomatous polyps as pre-cancerous lesions; patients were diagnosed using colonoscopy and pathology reports were available. Twenty control samples were also obtained from healthy subjects for comparison. RNA extraction and cDNA synthesis were followed by real-time PCR to evaluate lncRNA expression. The up-regulation of ANRIL in 20% of samples taken from polyp patients, combined with up-regulation in 65% of patients with CRC, confirmed the potential usefulness of ANRIL as a prognostic biomarker (AUC 0.95; P < 0.0001). BANCR relative expression analysis illustrated significant up-regulation in polyp (P < 0.04) and tumoural participants (P < 0.03) compared with normal control individuals. The expression patterns of ANRIL and BANCR in polyp cases were significantly correlated according to correlation analysis (r = 0.45, P < 0.045). ANRIL expression patterns in stool specimens of polyp and tumour cases supported the use of ANRIL as a prognostic biomarker for screening patients in the early stages of CRC. Up-regulation of BANCR in pre-cancerous lesions as well as down-regulation of ANRIL may also be a specific marker pair for easy, convenient and fast CRC prognosis.
长链非编码rna已被提出作为检测、预防和筛选各种恶性肿瘤的生物标志物。本研究通过粪便标本检测,评估了两种lncrna (ANRIL和BANCR)作为生物标志物在结直肠癌(CRC)早期检测中的应用,这是一种无创且具有成本效益的方法。从结直肠癌或腺瘤性息肉作为癌前病变转诊到医院的患者中共收集了40份粪便样本;患者通过结肠镜检查和病理报告进行诊断。还从健康受试者中获得20个对照样本进行比较。RNA提取和cDNA合成后采用实时荧光定量PCR检测lncRNA的表达。20%息肉患者样本中ANRIL表达上调,65%结直肠癌患者样本中ANRIL表达上调,证实了ANRIL作为预后生物标志物的潜在用途(AUC 0.95;P < 0.0001)。BANCR相对表达分析显示,与正常对照组相比,息肉组(P < 0.04)和肿瘤组(P < 0.03)显著上调。经相关分析,ANRIL与BANCR在息肉病例中的表达模式有显著相关性(r = 0.45, P < 0.045)。ANRIL在息肉和肿瘤患者粪便标本中的表达模式支持ANRIL作为筛查早期结直肠癌患者的预后生物标志物。癌前病变中BANCR的上调和ANRIL的下调也可能是简便、方便、快速预测结直肠癌预后的特异性标志物对。
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引用次数: 1
Phylogeographic study using nuclear genome sequences of Asip to infer the origins of ventral fur color variation in the house mouse Mus musculus. 利用Asip核基因组序列推断家鼠腹侧毛色变异起源的系统发育地理学研究。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-03-11 DOI: 10.1266/ggs.21-00075
Toki Takeishi, K. Fujiwara, N. Osada, Akihiko Mita, T. Takada, T. Shiroishi, Hitoshi Suzuki
While the house mouse (Mus musculus), widely distributed in Eurasia, is known to have substantial coat color variation between and within local populations, in both primary and secondary distribution areas, including the Japanese archipelago, the evolutionary history of the color variation is poorly understood. To address the ventral fur color variation, we quantified the lightness of museum skin specimens, and found that the southern subspecies, M. m. castaneus (CAS), has high and low lightness in dry and rainy geographic regions, respectively. The northern subspecies, M. m. musculus (MUS), has low and high levels of lightness in the high and middle latitudes of northern Eurasia, respectively. We examined sequence variation of the agouti signaling protein gene (Asip), which is known to be responsible for the ventral fur color. We performed phylogenetic analyses with 196 haplotype sequences of Asip (~180 kb) generated by phasing the whole-genome data of 98 wild mice reported previously. Network and phylogenetic tree construction revealed clustering of haplotypes representing the two subspecies, MUS and CAS. A number of subclusters with geographic affinities appeared within the subspecies clusters, in which the essential results were consistent with those reconstructed with whole mitochondrial genome data, indicating that the phased haplotype genome sequences of the nuclear genome can be a useful tool for tracing the dispersal of geographical lineages. The results of phylogeographic analysis showed that CAS mice with darker ventral fur possessed similar Asip haplotypes across the geographic distribution, suggesting that these haplotypes are major causes of the historical introduction of Asip haplotypes for darker ventral fur in mice from northern India to the peripheral areas, including the Japanese archipelago. Similarly, MUS in East Asia, which has a white abdomen, formed an Asip haplogroup with that from northern Iran, also with a white abdomen.
家鼠广泛分布在欧亚大陆,在包括日本群岛在内的一级和二级分布区,家鼠的毛色在当地种群之间和内部都有很大的变化,但人们对这种颜色变化的进化史知之甚少。为了解决腹侧皮毛颜色的变化,我们量化了博物馆皮肤标本的亮度,发现南方亚种M.castaneus(CAS)在干旱和多雨的地理区域分别具有高亮度和低亮度。北方亚种M.M.musculus(MUS)在欧亚大陆北部的高纬度和中纬度分别具有低亮度和高亮度。我们检测了agouti信号蛋白基因(Asip)的序列变异,已知该基因与腹侧毛色有关。我们对之前报道的98只野生小鼠的全基因组数据进行了分阶段分析,产生了196个Asip单倍型序列(~180kb)。网络和系统发育树的构建揭示了代表MUS和CAS两个亚种的单倍型的聚类。亚种群中出现了许多具有地理亲缘关系的亚群,其中基本结果与用线粒体全基因组数据重建的结果一致,表明核基因组的阶段性单倍型基因组序列可以成为追踪地理谱系分布的有用工具。系统地理学分析结果表明,具有深色腹毛的CAS小鼠在整个地理分布中具有相似的Asip单倍型,这表明这些单倍型是历史上从印度北部向包括日本群岛在内的周边地区引入深色腹毛Asip单倍型的主要原因。同样,东亚的MUS有一个白色的腹部,与伊朗北部的MUS形成了一个同样有白色腹部的Asip单倍群。
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引用次数: 0
A new allele of engrailed, enNK14, causes supernumerary spermathecae in Drosophila melanogaster. 在黑腹果蝇中,一个新的enk14等位基因导致了多精。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-03-09 DOI: 10.1266/ggs.21-00030
Yasuko Kato, Akiko Sawada, Kazuki Tonai, Hisashi Tatsuno, T. Uenoyama, M. Itoh
A spontaneous mutation, enNK14, was a new allele of engrailed (en) in Drosophila melanogaster. Females of enNK14 have three spermathecae, instead of two in wild type, under a wide range of developmental temperatures, while the males show no abnormal phenotype. Spermathecae of the mutant female can accept inseminated sperms, albeit with a delay of at least an hour until full acceptance compared with wild type. The time course of decrease in the number of stored sperms was thoroughly similar between the mutant and wild type. enNK14 females produced fewer progeny than wild type females despite storing a larger number of sperms. The delay of sperm entry and lower fecundity suggested some functional defects in secretory products of the spermathecae. In addition, some spermathecae in the mutant were accompanied by a mass of brown pigments in the adipose tissue surrounding the capsule. Six contiguous amino acids, Ser340-Ala345, were replaced by one Thr in enNK14. In another mutant, enspt, Ser325 was also shown to be substituted by a Cys. These amino acid changes were located within a serine-rich region, in which Ser325, Ser340 and Thr341 were suggested as targets of Protein Kinase C by an in silico analysis. The splicing pattern of en mRNA did not differ between enNK14 and wild type in embryo, larva, pupa or adult. Our results suggest that en plays an important role in determining the number of spermathecae as well as in sperm storage function in the Drosophila female.
一个自发突变,enNK14,是黑腹果蝇的一个新的等位基因。enNK14的雌性在不同的发育温度下有三个受精囊,而不是野生型的两个,而雄性没有表现出异常表型。突变雌性的精子库可以接受受精的精子,尽管与野生型相比,完全接受精子至少要延迟一个小时。突变体和野生型精子数量减少的时间过程完全相似。enNK14雌性比野生型雌性产生更少的后代,尽管储存了大量的精子。精子进入的延迟和繁殖力的降低表明受精囊分泌产物存在一些功能缺陷。此外,突变体中的一些受精囊在包膜周围的脂肪组织中伴有大量棕色色素。enNK14中的六个相邻氨基酸Ser340-Ala345被一个Thr取代。在另一个突变体enspt中,Ser325也被Cys取代。这些氨基酸变化位于富含丝氨酸的区域内,其中Ser325、Ser340和Thr341通过计算机分析被认为是蛋白激酶C的靶标。enNK14与野生型在胚胎、幼虫、蛹和成虫中的en mRNA剪接模式没有差异。我们的研究结果表明,en在决定果蝇雌性受精囊数量和精子储存功能方面发挥着重要作用。
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引用次数: 0
Development and evaluation of a loop-mediated isothermal amplification (LAMP) assay for quick identification of the Japanese salamander Hynobius tokyoensis. 环介导等温扩增(LAMP)快速鉴定东京火蜥蜴方法的建立与评价。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-02-23 Epub Date: 2021-11-07 DOI: 10.1266/ggs.21-00046
Tomoya Suzuki, Kanto Nishikawa, Yukuto Sato, Mamoru Toda

Species identification using molecular techniques has recently become common for various taxa. Loop-mediated isothermal amplification (LAMP) is one of the easiest and least expensive molecular identification methods. Although few studies have developed LAMP assays for amphibians, we believe that LAMP is also useful for identifying endangered amphibians. Hynobius tokyoensis and H. lichenatus occur in Honshu, Japan, and have parapatric distributions. They are similar morphologically, especially at early developmental stages, including eggs and larvae. Hynobius tokyoensis has been listed as a national endangered species in Japan since 2020, and unambiguous identification of these species is therefore important for their conservation and management. In this study, we developed a LAMP primer set for the mitochondrial cytochrome b region to detect H. tokyoensis, and we evaluated the LAMP assay using total genomic DNA from four H. tokyoensis and three H. lichenatus individuals from across most of their ranges. Our LAMP primer set could distinguish these two species. This study should help to establish LAMP assays for other endangered species and morphologically similar species.

近年来,利用分子技术进行物种鉴定已成为各种分类群的常用方法。环介导等温扩增(LAMP)是最简单、最便宜的分子鉴定方法之一。虽然很少有研究开发两栖动物的LAMP测定方法,但我们认为LAMP对识别濒危两栖动物也很有用。东京海狸鼠和地衣海狸鼠分布于日本本州,呈准巢分布。它们在形态上是相似的,特别是在早期发育阶段,包括卵和幼虫。自2020年以来,东京水蚤已被列为日本国家濒危物种,因此对这些物种的明确鉴定对其保护和管理至关重要。在这项研究中,我们开发了一套用于线粒体细胞色素b区域的LAMP引物来检测东京按蚊,并使用4个东京按蚊和3个地衣按蚊的基因组DNA对LAMP检测进行了评估。我们的LAMP引物可以区分这两个物种。本研究将有助于建立其他濒危物种和形态相似物种的LAMP检测方法。
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引用次数: 1
Genetic diversity and population structure of the long-tailed hamster Cricetulus longicaudatus in Shanxi Province, China. 山西省长尾仓鼠遗传多样性及种群结构研究
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-02-23 Epub Date: 2022-01-09 DOI: 10.1266/ggs.20-00060
Xin'gen Yang, Tinglin Wang, Hongfang Guo, Jing Yang, Bo Zou, Jianzhen Zhang

The long-tailed hamster Cricetulus longicaudatus is a dominant rodent in farmland of Shanxi Province, China, but little is known about its genetic diversity and population structure. In this study, the genomic DNAs of individuals from 13 populations captured in different fields of Shanxi were extracted and amplified by six pairs of microsatellite primers and by universal primers for mtDNA COI gene sequences. Our data revealed significant departure from Hardy-Weinberg equilibrium in four of the 13 populations. In all 13 populations, the mean observed heterozygosity was significantly lower than the expected heterozygosity. Meanwhile, the mean inbreeding coefficient was statistically significant, which indicated non-random mating within populations. The pairwise genetic distance and natural logarithm of linear geographical distance were not significantly correlated for any C. longicaudatus populations. However, the correlation between genetic distance and resistance distance based on mountain landscape was significant, suggesting that the mountain landscape is an important factor affecting gene flow of C. longicaudatus. Pairwise FST analysis of population structure showed moderate to high genetic differentiation among populations, and all individuals could be divided into two gene clusters. Phylogenetic analysis based on COI sequences also showed that many individuals originated from a single haplotype, suggesting the existence of gene exchange among these populations at some time in the past. Our research should provide a scientific basis for the analysis of genetic differentiation and gene flow among populations of C. longicaudatus.

长尾仓鼠是山西农田啮齿动物的优势种,但对其遗传多样性和种群结构了解甚少。本研究利用6对微卫星引物和mtDNA COI基因序列的通用引物,对在山西不同地区捕获的13个种群的个体进行基因组dna提取和扩增。我们的数据显示13个种群中有4个明显偏离Hardy-Weinberg平衡。在所有13个群体中,观察到的平均杂合度显著低于预期的杂合度。同时,平均近交系数有统计学意义,表明种群内交配是非随机的。各居群间的遗传距离与线性地理距离的自然对数均无显著相关。然而,基于山地景观的遗传距离与抗性距离的相关性显著,表明山地景观是影响长尾藤基因流动的重要因素。群体结构两两FST分析显示群体间存在中等至高度的遗传分化,所有个体均可划分为两个基因簇。基于COI序列的系统发育分析也表明,许多个体起源于单一单倍型,表明这些群体之间在过去的某个时间存在基因交换。本研究为分析长尾竹居群间的遗传分化和基因流动提供了科学依据。
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引用次数: 0
Silencing and anti-silencing mechanisms that shape the epigenome in plants. 形成植物表观基因组的沉默和反沉默机制。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-02-23 Epub Date: 2021-10-29 DOI: 10.1266/ggs.21-00041
Soichi Inagaki

Epigenome information mediates genome function and maintenance by regulating gene expression and chromatin organization. Because the epigenome pattern can change in response to internal and external environments, it may underlie an adaptive genome response that modulates phenotypes during development and in changing environments. Here I summarize recent progress in our understanding of how epigenome patterns are shaped and modulated by concerted actions of silencing and anti-silencing factors mainly in Arabidopsis thaliana. I discuss the dynamic nature of epigenome regulation, which is realized by cooperation and counteraction among silencing and anti-silencing factors, and how the dynamic epigenome mediates robust and plastic responses of plants to fluctuating environments.

表观基因组信息通过调节基因表达和染色质组织介导基因组功能和维持。由于表观基因组模式可以响应内部和外部环境而改变,因此它可能是适应性基因组反应的基础,在发育过程中和不断变化的环境中调节表型。在这里,我总结了我们对拟南芥中沉默和抗沉默因子如何形成和调节表观基因组模式的理解的最新进展。我讨论了表观基因组调控的动态性,这是通过沉默和反沉默因子之间的合作和相互作用来实现的,以及动态表观基因组如何介导植物对波动环境的稳健和可塑性反应。
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引用次数: 2
Development and characterization of microsatellite markers for Rhododendron purdomii (Ericaceae) using next-generation sequencing. 利用新一代测序技术开发和鉴定杜鹃花微卫星标记。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-02-23 Epub Date: 2021-12-15 DOI: 10.1266/ggs.21-00031
Ningning Zhang, Mengyun Qin, Shixin Zhu, Ziyang Huang, Hao Dong, Yang Yang, Lili Yang, Yang Lu

Rhododendron purdomii (Ericaceae) is an endangered ornamental species endemic to the Qinling Mountains of China. Due to the impact of climate change and human disturbance, R. purdomii is threatened by habitat loss, and conservation of this species is urgently needed. In this study, we developed and characterized 13 novel microsatellite markers for R. purdomii based on next-generation sequencing data. For the 13 microsatellite markers in three R. purdomii populations, the number of alleles ranged from two to 12, the number of effective alleles was from 1.000 to 8.892, Shannon's information index was from 0.000 to 2.320, and the observed and expected heterozygosity were from 0.000 to 1.000 and from 0.000 to 0.888, respectively. Private alleles were found in all three populations. Moderate differentiation between population pairs was indicated by pairwise FST values. The microsatellite markers developed in this study will provide opportunities for examining the genetic diversity and population structure of R. purdomii and contribute to the effective conservation of this species.

杜鹃花(杜鹃花科)是中国秦岭特有的濒危观赏植物。由于气候变化和人为干扰的影响,白桦生境受到严重威胁,亟待保护。在这项研究中,我们基于新一代测序数据开发并鉴定了13个新的purdomii微卫星标记。13个微卫星标记的等位基因数为2 ~ 12个,有效等位基因数为1.000 ~ 8.892个,香农信息指数为0.000 ~ 2.320,观察杂合度为0.000 ~ 1.000,期望杂合度为0.000 ~ 0.888。在所有三个人群中均发现了私有等位基因。成对FST值表明种群对之间存在中度分化。本研究所建立的微卫星标记将为深入研究紫金石的遗传多样性和种群结构提供机会,并为该物种的有效保护做出贡献。
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引用次数: 2
Heat memory in plants: histone modifications, nucleosome positioning and miRNA accumulation alter heat memory gene expression. 植物的热记忆:组蛋白修饰、核小体定位和miRNA积累改变热记忆基因的表达。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-02-23 Epub Date: 2021-09-16 DOI: 10.1266/ggs.21-00040
Nobutoshi Yamaguchi

Plant adaptation to high temperature, often referred to as heat acclimation, is a process in which exposure to moderately high temperatures increases a plant's tolerance to subsequent (normally) lethal high temperatures. Plants store heat experience information (heat memory) obtained from previous exposure to high temperatures for several days and develop future temperature responsiveness. However, our understanding of heat acclimation is very limited. In the model plant Arabidopsis thaliana, changes in the expression patterns of heat memory genes play a central role in regulating plant survival and adaptation to recurring heat stress. Heat stress-related transcription factors and histone-modifying enzymes function in the sensitized expression of heat memory genes via the deposition and removal of histone modifications. Chromatin-remodeling complexes and miRNA accumulation also trigger the sustained expression of heat memory genes. In this review, I describe studies of heat acclimation that have provided important insights into the molecular mechanisms that lead to flexible and reversible gene expression upon heat stress in plants.

植物对高温的适应,通常被称为热驯化,是一个暴露在中等高温下增加植物对随后(通常)致命高温的耐受性的过程。植物可以储存从高温环境中获得的热经验信息(热记忆)数天,并发展出未来的温度响应能力。然而,我们对热驯化的认识非常有限。在模式植物拟南芥(Arabidopsis thaliana)中,热记忆基因表达模式的变化在调节植物生存和适应反复热胁迫中起着核心作用。热应激相关转录因子和组蛋白修饰酶通过沉积和去除组蛋白修饰在热记忆基因的敏化表达中起作用。染色质重塑复合物和miRNA积累也触发热记忆基因的持续表达。在这篇综述中,我描述了热驯化的研究,这些研究为植物热胁迫下导致灵活和可逆的基因表达的分子机制提供了重要的见解。
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引用次数: 6
Spatiotemporal regulation and roles of reproductive phasiRNAs in plants. 植物生殖相rna的时空调控及其作用。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2022-02-23 Epub Date: 2021-11-11 DOI: 10.1266/ggs.21-00042
Reina Komiya
Since co-suppression was discovered as a pioneer silencing phenomenon of RNA interference (RNAi) in petunia in 1990, many types of small RNAs have been identified in the RNAi pathway among various eukaryotes. In plants, a large number of 21- or 24-nucleotide (nt) phased small interfering RNAs (phasiRNAs) are produced via processing of long RNA precursors by Dicer-like proteins. However, the roles of phasiRNAs remain largely unknown. The development of imaging technology and RNA profiling has clarified the spatiotemporal regulation of phasiRNAs, and subsequently the different functions of 21-nt trans-acting phasiRNAs and 24-nt cis-regulatory phasiRNAs during male organ development. This review focuses on the biogenesis, diversification, spatiotemporal expression pattern and function of phasiRNAs in plants.
自1990年在矮牵牛中发现共抑制作为RNA干扰(RNAi)沉默的先驱现象以来,在各种真核生物的RNAi途径中已经鉴定出许多类型的小RNA。在植物中,dicer样蛋白通过加工长RNA前体产生大量的21或24核苷酸相小干扰RNA (phasiRNAs)。然而,phasirna的作用在很大程度上仍然未知。随着成像技术和RNA谱的发展,phasiRNAs的时空调控得以阐明,并由此揭示了21-nt反式作用phasiRNAs和24-nt顺式调节phasiRNAs在男性器官发育过程中的不同功能。本文就植物相rna的发生、多样性、时空表达模式及功能等方面的研究进展作一综述。
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引用次数: 4
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