scholarly journals Plant Extracellular Vesicles Contain Diverse Small RNA Species and Are Enriched in 10- to 17-Nucleotide “Tiny” RNAs

2019 ◽  
Vol 31 (2) ◽  
pp. 315-324 ◽  
Author(s):  
Patricia Baldrich ◽  
Brian D. Rutter ◽  
Hana Zand Karimi ◽  
Ram Podicheti ◽  
Blake C. Meyers ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Small Rna ◽  
Small Rna Species

scholarly journals Mining diverse small RNA species in the deep transcriptome

2015 ◽  
Vol 40 (1) ◽  
pp. 4-7 ◽  
Author(s):  
Kasey C. Vickers ◽  
Leslie A. Roteta ◽  
Holli Hucheson-Dilks ◽  
Leng Han ◽  
Yan Guo
Keyword(s):  
Small Rna ◽  
Small Rna Species

Synthesis of two classes of small RNA species in vivo and in vitro

Biochemistry ◽  
1977 ◽  
Vol 16 (20) ◽  
pp. 4520-4525 ◽  
Author(s):  
Gary Zieve ◽  
Bernd Joachim Benecke ◽  
Sheldon Penman
Keyword(s):  
Small Rna ◽  
Small Rna Species

scholarly journals 3D cell culture stimulates the secretion of in vivo like extracellular vesicles

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Sirisha Thippabhotla ◽  
Cuncong Zhong ◽  
Mei He
Keyword(s):  
Cervical Cancer ◽  
Cell Culture ◽  
Cancer Patient ◽  
Extracellular Vesicles ◽  
Small Rna ◽  
Small Rnas ◽  
3D Culture ◽  
Cervical Cancer Patient ◽  
Tumor Biomarker

Abstract For studying cellular communications ex-vivo, a two-dimensional (2D) cell culture model is currently used as the “gold standard”. 2D culture models are also widely used in the study of RNA expression profiles from tumor cells secreted extracellular vesicles (EVs) for tumor biomarker discovery. Although the 2D culture system is simple and easily accessible, the culture environment is unable to represent in vivo extracellular matrix (ECM) microenvironment. Our study observed that 2D- culture derived EVs showed significantly different profiles in terms of secretion dynamics and essential signaling molecular contents (RNAs and DNAs), when compared to the three-dimensional (3D) culture derived EVs. By performing small RNA next-generation sequencing (NGS) analysis of cervical cancer cells and their EVs compared with cervical cancer patient plasma EV-derived small RNAs, we observed that 3D- culture derived EV small RNAs differ from their parent cell small RNA profile which may indicate a specific sorting process. Most importantly, the 3D- culture derived EV small RNA profile exhibited a much higher similarity (~96%) to in vivo circulating EVs derived from cervical cancer patient plasma. However, 2D- culture derived EV small RNA profile correlated better with only their parent cells cultured in 2D. On the other hand, DNA sequencing analysis suggests that culture and growth conditions do not affect the genomic information carried by EV secretion. This work also suggests that tackling EV molecular alterations secreted into interstitial fluids can provide an alternative, non-invasive approach for investigating 3D tissue behaviors at the molecular precision. This work could serve as a foundation for building precise models employed in mimicking in vivo tissue system with EVs as the molecular indicators or transporters. Such models could be used for investigating tumor biomarkers, drug screening, and understanding tumor progression and metastasis.

Co-purification of a small RNA species with multicatalytic proteinase (proteasome) from rat liver

FEBS Letters ◽  
1991 ◽  
Vol 279 (2) ◽  
pp. 351-355 ◽  
Author(s):  
Helen E. Skilton ◽  
Ian C. Eperon ◽  
A.Jennifer Rivett
Keyword(s):  
Rat Liver ◽  
Small Rna ◽  
Multicatalytic Proteinase ◽  
Small Rna Species

scholarly journals Comparative profiling of small RNAs of pig seminal plasma and ejaculated and epididymal sperm

Reproduction ◽  
2017 ◽  
Vol 153 (6) ◽  
pp. 785-796 ◽  
Author(s):  
Cai Chen ◽  
Han Wu ◽  
Dan Shen ◽  
Saisai Wang ◽  
Li Zhang ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Small Rna ◽  
Small Rnas ◽  
Target Genes ◽  
Large Population ◽  
Small Nuclear Rna ◽  
Epididymal Sperm ◽  
Zygote Development ◽  
Similarities And Differences ◽  
Small Rna Species

The similarities and differences of small RNAs in seminal plasma, epididymal sperm and ejaculated sperm remain largely undefined. We conducted a systematic comparative analysis of small RNA profiles in pig ejaculated sperm, epididymal sperm and seminal plasma and found that the diversity distribution of small RNA species was generally similar, whereas the abundance of small RNAs is dramatically different across the three libraries; miRNAs and small RNAs derived from rRNA, tRNA, small nuclear RNA, 7SK RNA, NRON RNA and cis-regulatory RNA were enriched in the three libraries, but piRNA was absent. A large population of small RNAs from ejaculated sperm are ejaculated sperm specific, and only 8–30% of small RNAs overlapped with those of epididymal sperm or seminal plasma and a small proportion (5–18%) of small RNAs were shared in the three libraries, suggesting that, in addition to the testes, sperm RNAs may also originate from seminal plasma, epididymis as well as other resources. Most miRNAs were co-distributed but differentially expressed across the three libraries, with epididymal sperm exhibiting the highest abundance, followed by ejaculated sperm and seminal plasma. The prediction of target genes of the top 10 highly expressed miRNAs across the three libraries revealed that these miRNAs may be involved in spermatogenesis, zygote development and the interaction between the environment and animals. Our study provides the first description of the similarities and differences of small RNA profiles in ejaculated sperm, epididymal sperm and seminal plasma and indicates that sperm RNA may have origins other than the testes.

scholarly journals Characterization of the small RNA content of Trypanosoma cruzi extracellular vesicles

2014 ◽  
Vol 193 (2) ◽  
pp. 71-74 ◽  
Author(s):  
Ethel Bayer-Santos ◽  
Fábio Mitsuo Lima ◽  
Jeronimo Conceição Ruiz ◽  
Igor C. Almeida ◽  
José Franco da Silveira
Keyword(s):  
Trypanosoma Cruzi ◽  
Extracellular Vesicles ◽  
Small Rna ◽  
Rna Content

scholarly journals Relationships of Alzheimer’s disease and apolipoprotein E genotypes with small RNA and protein cargo of brain tissue extracellular vesicles

2020 ◽  
Author(s):  
Yiyao Huang ◽  
Tom A. P. Driedonks ◽  
Lesley Cheng ◽  
Andrey Turchinovich ◽  
Harinda Rajapaksha ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Extracellular Vesicles ◽  
Small Rna ◽  
Apoe Genotype ◽  
Tau Proteins ◽  
Small Rna Sequencing ◽  
Health Crisis ◽  
Metabolic Functions ◽  
Apolipoprotein E Genotypes

ABSTRACTAlzheimer’s disease (AD) is a public health crisis that grows as populations age. Hallmarks of this neurodegenerative disease include aggregation of beta-amyloid peptides and hyperphosphorylated tau proteins in the brain. Variants of the APOE gene are the greatest known risk factors for sporadic AD. As emerging players in AD pathophysiology, extracellular vesicles (EVs) contain proteins, lipids, and RNAs and are involved in disposal of cellular toxins and intercellular communication. AD-related changes in the molecular composition of EVs may contribute to pathophysiology and lend insights into disease mechanisms. We recently adapted a method for separation of brain-derived EVs (bdEVs) from post-mortem tissues. Using this method, we isolated bdEVs from AD patients with different APOE genotypes (n=23) and controls (n=7). bdEVs were counted, sized, and subjected to parallel small RNA sequencing and proteomic analysis. Numerous bdEV-associated RNAs and proteins correlated with AD pathology and APOE genotype. Some of the identified entities have been implicated previously in important AD-related pathways, including amyloid processing, neurodegeneration, and metabolic functions. These findings provide further evidence that bdEVs and their molecular cargo modulate development and progression of AD.

scholarly journals Functional analysis of virus and host components that mediate potyvirus-induced diseases

2008 ◽  
Author(s):  
Steven A. Whitham ◽  
Amit Gal-On ◽  
Tzahi Arazi
Keyword(s):  
Small Rna ◽  
Small Rnas ◽  
Binding Activity ◽  
Symptom Expression ◽  
Wild Type ◽  
Disease Symptoms ◽  
Virus Accumulation ◽  
Plant Genes ◽  
Small Rna Species ◽  
Host Genes

The mechanisms underlying the development of symptoms in response to virus infection remain to be discovered in plants. Insight into symptoms induced by potyviruses comes from evidence implicating the potyviral HC-Pro protein in symptom development. In particular, recent studies link the development of symptoms in infected plants to HC-Pro's ability to interfere with small RNA metabolism and function in plant hosts. Moreover, mutation of the highly conserved FRNK amino acid motif to FINK in the HC-Pro of Zucchini yellow mosaic virus (ZYMV) converts a severe strain into an asymptomatic strain, but does not affect virus accumulation in cucurbit hosts. The ability of this FINK mutation to uncouple symptoms from virus accumulation creates a unique opportunity to study symptom etiology, which is usually confounded by simultaneous attenuation of both symptoms and virus accumulation. Our goal was to determine how mutations in the conserved FRNK motif affect host responses to potyvirus infection in cucurbits and Arabidopsis thaliana. Our first objective was to define those amino acids in the FRNK motif that are required for symptoms by mutating the FRNK motif in ZYMV and Turnip mosaic virus (TuMV). Symptom expression and accumulation of resulting mutant viruses in cucurbits and Arabidopsis was determined. Our second objective was to identify plant genes associated with virus disease symptoms by profiling gene expression in cucurbits and Arabidopsis in response to mutant and wild type ZYMV and TuMV, respectively. Genes from the two host species that are differentially expressed led us to focus on a subset of genes that are expected to be involved in symptom expression. Our third objective was to determine the functions of small RNA species in response to mutant and wild type HC-Pro protein expression by monitoring the accumulation of small RNAs and their targets in Arabidopsis and cucurbit plants infected with wild type and mutant TuMV and ZYMV, respectively. We have found that the maintenance of the charge of the amino acids in the FRNK motif of HC-Pro is required for symptom expression. Reduced charge (FRNA, FRNL) lessen virus symptoms, and maintain the suppression of RNA silencing. The FRNK motif is involved in binding of small RNA species including microRNAs (miRNA) and short interfering RNAs (siRNA). This binding activity mediated by the FRNK motif has a role in protecting the viral genome from degradation by the host RNA silencing system. However, it also provides a mechanism by which the FRNK motif participates in inducing the symptoms of viral infection. Small RNA species, such as miRNA and siRNA, can regulate the functions of plant genes that affect plant growth and development. Thus, this binding activity suggests a mechanism by which ZYMVHC-Pro can interfere with plant development resulting in disease symptoms. Because the host genes regulated by small RNAs are known, we have identified candidate host genes that are expected to play a role in symptoms when their regulation is disrupted during viral infections. As a result of this work, we have a better understanding of the FRNK amino acid motif of HC-Pro and its contribution to the functions of HC-Pro, and we have identified plant genes that potentially contribute to symptoms of virus infected plants when their expression becomes misregulated during potyviral infections. The results set the stage to establish the roles of specific host genes in viral pathogenicity. The potential benefits include the development of novel strategies for controlling diseases caused by viruses, methods to ensure stable expression of transgenes in genetically improved crops, and improved potyvirus vectors for expression of proteins or peptides in plants.   

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