猪miR-199a-5p对脂质生成的影响及其靶基因的预测和分析

本研究旨在分析miR-199a-5p对猪肌内脂肪细胞脂质生成的影响及作用机制。采集淮南猪不同育肥时期(育肥前期、中期和后期)的背最长肌和皮下脂肪组织,通过实时荧光定量PCR分析其表达变化趋势;合成miR-199a-5p的mimics,转染猪原代肌内脂肪细胞,诱导分化后通过油红O染色观察过表达miR-199a-5p对脂质生成的影响;结合之前筛选到的脂肪型和瘦肉型猪差异表达mRNAs、lncRNAs和circRNAs,使用miRanda软件筛选miR-199a-5p的靶分子,并用GO和KEGG富集分析法对这些mRNAs、lncRNAs的共表达基因和circRNAs的来源基因进行功能分析。结果显示,随着育肥进程,miR-199a-5p在背最长肌中表达量持续上调,而在皮下脂肪组织中表达量先下调后上调;过表达miR-199a-5p可抑制肌内脂肪细胞的脂质生成。共筛选到9个mRNAs、5个lncRNAs和1258个circRNAs包含miR-199a-5p结合位点,GO分析主要富集于内质网和肌动蛋白结合,KEGG分析发现主要富集于糖、脂质和蛋白质代谢。提示miR-199a-5p可能通过与lncRNAs和circRNAs互作间接调控靶基因,参与调控肌肉发育、脂质生成和代谢,可作为影响肉质性状的候选miRNA。     

猪miR-199a-5p对脂质生成的影响及其靶基因的预测和分析

本研究旨在分析miR-199a-5p对猪肌内脂肪细胞脂质生成的影响及作用机制。采集淮南猪不同育肥时期（育肥前期、中期和后期）的背最长肌和皮下脂肪组织,通过实时荧光定量PCR分析其表达变化趋势;合成miR-199a-5p的mimics,转染猪原代肌内脂肪细胞,诱导分化后通过油红O染色观察过表达miR-199a-5p对脂质生成的影响;结合之前筛选到的脂肪型和瘦肉型猪差异表达mRNAs、lncRNAs和circRNAs,使用miRanda软件筛选miR-199a-5p的靶分子,并用GO和KEGG富集分析法对这些mRNAs、lncRNAs的共表达基因和circRNAs的来源基因进行功能分析。结果显示,随着育肥进程,miR-199a-5p在背最长肌中表达量持续上调,而在皮下脂肪组织中表达量先下调后上调;过表达miR-199a-5p可抑制肌内脂肪细胞的脂质生成。共筛选到9个mRNAs、5个lncRNAs和1 258个circRNAs包含miR-199a-5p结合位点,GO分析主要富集于内质网和肌动蛋白结合,KEGG分析发现主要富集于糖、脂质和蛋白质代谢。提示miR-199a-5p可能通过与lncRNAs和circRNAs互作间接调控靶基因,参与调控肌肉发育、脂质生成和代谢,可作为影响肉质性状的候选miRNA。     

动植物环状RNA研究进展与展望

为促进植物环状RNA(Circular RNAs)的研究进程,利用文献综述法检索近三年(2018—2020)发表的环状RNA文献,从功能发挥、研究手段及应用前景等方面分别对动物和植物环状RNA的研究进展进行整理讨论。结果表明:环状RNA可作为竞争性内源RNA与microRNAs(MiRNAs)特异性结合,间接调控蛋白表达水平。其还可调节来源基因的转录效率、滚环式翻译多肽及作为蛋白支架调控细胞进程。截止目前,临床有关环状RNA的研究最为深入;经过潜在诊断、预断和治疗方法评估后,环状RNA有望成为诊疗疾病的生物标志物。而植物环状RNA在不同生长发育阶段、激素刺激及逆境胁迫等条件下也展现出丰富的差异表达模式,初步体现其功能的多样性。但与动物环状RNA的研究深度相比,植物环状RNA研究还处于兴起阶段,与植物环状RNA相关的数据库和分析工具还有待开发。综上,本文重点评述动植物体内不同类型环状RNA在生成机制、剪切识别位点、功能发挥及应用潜力上的特异性和同源性,讨论动植物环状RNA研究存在的主要问题并对深入开展植物环状RNA研究进行展望。     

捻转血矛线虫耐药虫株与敏感虫株circRNAs差异表达分析

为分析捻转血矛线虫伊维菌素敏感虫株与耐药虫株之间差异表达的circRNAs在调控耐药性中的作用,本研究以捻转血矛线虫敏感虫株和耐药虫株为研究对象,采用Illumina Hiseq 2 000平台进行高通量测序及cDNA文库的构建,利用相关生物信息学软件对测得的circRNAs进行差异性分析,同时对来源基因进行GO和KEGG pathway功能富集。运用TargetScan和Miranda软件预测关键circRNAs靶向的miRNAs,探究circRNAs-miRNAs的作用。随后又随机选取5个显著差异的circRNAs,进行RT-qPCR检测并与转录组测序结果进行一致性评定。结果显示：1）敏感虫株与耐药虫株共筛选出677个差异表达的circRNAs,其中47个显著差异,23个显著上调,24个显著下调。2）所有差异circRNAs的来源基因GO富集分析显示,共富集到生物过程336个、细胞组分59个和分子功能110个,主要与代谢过程、细胞膜结构和催化活性等相关;KEGG富集分析显示,富集到257条通路,主要与代谢（脂肪酸生物合成、甘油磷脂代谢和丙酸代谢）、胰高血糖素信号通路和AMPK信号通...     

犬弓首蛔虫感染比格犬不同阶段肝circRNAs表达模式的分析

旨在探究终末宿主肝环状RNA （circRNAs）在犬弓首蛔虫诱导致病过程中的潜在调控作用。18只6~7周龄的比格犬被平均分为3组（感染后0.5 d组、感染后1 d组和感染后36 d组），每组包括感染和对照各3只，分别于感染后对应的时间点收集肝样品。提取感染和对照组肝的总RNA，利用高通量RNA测序，构建circRNA文库，对差异表达的circRNAs进行分析，并对差异表达circRNAs的亲本基因进行GO注释和KEGG富集分析。随机选取9个差异表达circRNAs进行实时荧光定量PCR验证。结果显示，与对照组相比，在感染后0.5、1和36 d分别有94、103和84个犬肝的差异表达circRNAs。Venn图显示，3个感染阶段没有共同表达的差异circRNAs；GO和KEGG分析显示，差异表达的circRNAs参与了宿主肝免疫与炎症的相关通路，其中差异表达的circRNA novel_circ_0016108、novel_circ_0016184和novel_circ_0027468与犬弓首蛔虫引起的天然免疫相关；novel_circ_0002212参与了犬弓首蛔虫引起的肝炎症反应；novel_circ_0002212和novel_circ_0019907在宿主肝抵抗犬弓首蛔虫侵袭的过程中发挥作用。RT-qPCR验证结果显示大多数差异表达的circRNAs与高通量测序结果一致。结果表明，肝circRNAs在犬弓首蛔虫感染终末宿主的过程中发挥着重要作用，这为进一步探究犬弓首蛔虫与宿主之间的相互作用提供了新的信息，并且也为促进疾病干预措施的制定提供参考。     

Biological functions of circRNAs and their progress in livestock and poultry

CircRNAs are a new member of endogenous RNAs, which characterized by a closed-loop structure without terminated 5′ caps and 3′ tails via covalent bonding, which are evolutionarily conserved among different species and often exhibit tissue or developmental stage-specific expression. So far, thousands of circRNAs have been discovered in eukaryotic cells. CircRNAs are more stable due to its resistance to exonuclease implying important biological functions in all kingdoms of life. They could function as miRNA sponges, interfere with splicing and bind to protein to regulate the expression of host genes and so on. In addition, emerging evidence suggests that circRNAs are closely related to a series of physiological processes in livestock and poultry. In this review, we summarized the biogenesis mechanism, major biological function and detection methods and focused on research advance of circRNAs in livestock and poultry, aiming at providing certain reference value and novel techniques for the development of new molecular genetic markers and breeding of livestock and poultry. Meanwhile, we hope this review could show significant prospect for other researches.     

circRNAs: post-transcription regulation in cerebrovascular diseases

Circular RNAs (circRNAs) are a type of covalently closed circular non-coding RNAs that are expressed across varieties of life forms. CircRNAs have a variety of characteristics such as stable structures, tissue-specific expression and the function of microRNA sponges in the regulation of genes. CircRNAs are abundant, highly expressed in neural tissues, emerging evidence also indicates that circRNAs play an important role in the pathophysiologic process of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis and other neurological diseases. As one of the most important neurological diseases, the pathogenesis of brain damage and repair after ischemia has not yet been evaluated, and stroke related circRNAs are of great significance. In this review, we summarize related regulations, and the potential regulatory mechanisms of cicrcRNAs in cerebrovascular diseases, and the important role of circRNAs in post stroke pathophysiologic process is also discussed. CircRNAs are expected to be a new drug target, and provide new theoretical basis for the diagnosis and treatment of cerebrovascular diseases.