Secondarysmallinterfering RNA (secondary siRNA)

2015 ◽  
pp. 1-1
Keyword(s):  
Secondary Sirna

scholarly journals Lack of pairing during meiosis triggers multigenerational transgene silencing in Caenorhabditis elegans

2015 ◽  
Vol 112 (20) ◽  
pp. E2667-E2676 ◽  
Author(s):  
Luciana E. Leopold ◽  
Bree N. Heestand ◽  
Soobin Seong ◽  
Ludmila Shtessel ◽  
Shawn Ahmed
Keyword(s):  
Caenorhabditis Elegans ◽  
Small Rna ◽  
Transgene Expression ◽  
Single Copy ◽  
Transgene Silencing ◽  
Meiotic Pairing ◽  
Homologous Chromosomes ◽  
Diverse Species ◽  
Secondary Sirna ◽  
Interfering Rna

Single-copy transgenes in Caenorhabditis elegans can be subjected to a potent, irreversible silencing process termed small RNA-induced epigenetic silencing (RNAe). RNAe is promoted by the Piwi Argonaute protein PRG-1 and associated Piwi-interacting RNAs (piRNAs), as well as by proteins that promote and respond to secondary small interfering RNA (siRNA) production. Here we define a related siRNA-mediated silencing process, termed “multigenerational RNAe,” which can occur for transgenes that are maintained in a hemizygous state for several generations. We found that transgenes that contain either GFP or mCherry epitope tags can be silenced via multigenerational RNAe, whereas a transgene that possesses GFP and a perfect piRNA target site can be rapidly and permanently silenced via RNAe. Although previous studies have shown that PRG-1 is typically dispensable for maintenance of RNAe, we found that both initiation and maintenance of multigenerational RNAe requires PRG-1 and the secondary siRNA biogenesis protein RDE-2. Although silencing via RNAe is irreversible, we found that transgene expression can be restored when hemizygous transgenes that were silenced via multigenerational RNAe become homozygous. Furthermore, multigenerational RNAe was accelerated when meiotic pairing of the chromosome possessing the transgene was abolished. We propose that persistent lack of pairing during meiosis elicits a reversible multigenerational silencing response, which can lead to permanent transgene silencing. Multigenerational RNAe may be broadly relevant to single-copy transgenes used in experimental biology and to shaping the epigenomic landscape of diverse species, where genomic polymorphisms between homologous chromosomes commonly result in unpaired DNA during meiosis.

scholarly journals MicroRNAs from the parasitic plant Cuscuta campestris target host messenger RNAs

2017 ◽  
Author(s):  
Saima Shahid ◽  
Gunjune Kim ◽  
Nathan R. Johnson ◽  
Eric Wafula ◽  
Feng Wang ◽  
...  
Keyword(s):  
Host Plants ◽  
Messenger Rnas ◽  
Host Gene Expression ◽  
Mrna Accumulation ◽  
Directional Movement ◽  
Plant Mirna ◽  
Cuscuta Campestris ◽  
Target Sites ◽  
Secondary Sirna ◽  
Interfering Rna

First paragraphDodders (Cuscuta spp.) are obligate parasitic plants that obtain water and nutrients from the stems of host plants via specialized feeding structures called haustoria. Dodder haustoria facilitate bi-directional movement of viruses, proteins, and mRNAs between host and parasite1, but the functional effects of these movements are not clear. Here we show that C. campestris haustoria accumulate high levels of many novel microRNAs (miRNAs) while parasitizing Arabidopsis thaliana hosts. Many of these miRNAs are 22 nts long, a usually rare size of plant miRNA associated with amplification of target silencing through secondary small interfering RNA (siRNA) production2. Several A. thaliana mRNAs are targeted by C. campestris 22 nt miRNAs during parasitism, resulting in mRNA cleavage, secondary siRNA production, and decreased mRNA accumulation levels. Hosts with mutations in two of the targets supported significantly higher growth of C. campestris. Homologs of target mRNAs from diverse plants also have predicted target sites to induced C. campestris miRNAs, and the same miRNAs are expressed and active against host targets when C. campestris parasitizes a different host, Nicotiana benthamiana. These data show that C. campestris miRNAs act as trans-species regulators of host gene expression, and suggest that they may act as virulence factors during parasitism.

scholarly journals Genome-wide CRISPR screening identifies new regulators of glycoprotein secretion

2019 ◽  
Author(s):  
Stephanie J. Popa ◽  
Julien Villeneuve ◽  
Sarah Stewart ◽  
Esther Perez Garcia ◽  
Anna Petrunkina Harrison ◽  
...  
Keyword(s):  
Protein Secretion ◽  
Subcellular Localisation ◽  
Novel Genes ◽  
Genome Wide ◽  
Fundamental Process ◽  
Crispr Screen ◽  
Secondary Sirna ◽  
Golgi Structure ◽  
Glycoprotein Secretion ◽  
Selection Of

ABSTRACTThe fundamental process of protein secretion from eukaryotic cells has been well described for many years, yet gaps in our understanding of how this process is regulated remain. With the aim of identifying novel genes involved in the secretion of glycoproteins, we used a screening pipeline consisting of a pooled genome-wide CRISPR screen followed by secondary siRNA screening of the hits to identify and validate several novel regulators of protein secretion. We present approximately 50 novel genes not previously associated with protein secretion, many of which also had an effect on the structure of the Golgi apparatus. We further studied a small selection of hits to investigate their subcellular localisation. One of these, GPR161, is a novel Golgi-resident protein that we propose maintains Golgi structure via an interaction with golgin A5.

scholarly journals Genome-wide CRISPR screening identifies new regulators of glycoprotein secretion

2019 ◽  
Vol 4 ◽  
pp. 119
Author(s):  
Stephanie Popa ◽  
Julien Villeneuve ◽  
Sarah Stewart ◽  
Esther Perez Garcia ◽  
Anna Petrunkina Harrison ◽  
...  
Keyword(s):  
Protein Secretion ◽  
Subcellular Localisation ◽  
Novel Genes ◽  
Genome Wide ◽  
Fundamental Process ◽  
Crispr Screen ◽  
Secondary Sirna ◽  
Golgi Structure ◽  
Glycoprotein Secretion ◽  
Selection Of

Background: The fundamental process of protein secretion from eukaryotic cells has been well described for many years, yet gaps in our understanding of how this process is regulated remain. Methods: With the aim of identifying novel genes involved in the secretion of glycoproteins, we used a screening pipeline consisting of a pooled genome-wide CRISPR screen, followed by secondary siRNA screening of the hits to identify and validate several novel regulators of protein secretion. Results: We present approximately 50 novel genes not previously associated with protein secretion, many of which also had an effect on the structure of the Golgi apparatus. We further studied a small selection of hits to investigate their subcellular localisation. One of these, GPR161, is a novel Golgi-resident protein that we propose maintains Golgi structure via an interaction with golgin A5. Conclusions: This study has identified new factors for protein secretion involved in Golgi homeostasis.

scholarly journals Plant secondary siRNA production determined by microRNA-duplex structure

2012 ◽  
Vol 109 (7) ◽  
pp. 2461-2466 ◽  
Author(s):  
P. A. Manavella ◽  
D. Koenig ◽  
D. Weigel
Keyword(s):  
Duplex Structure ◽  
Secondary Sirna

Analyses of a Glycine max Degradome Library Identify microRNA Targets and MicroRNAs that Trigger Secondary SiRNA Biogenesis

2013 ◽  
Vol 55 (2) ◽  
pp. 160-176 ◽  
Author(s):  
Zheng Hu ◽  
Qiyan Jiang ◽  
Zhiyong Ni ◽  
Rui Chen ◽  
Shuo Xu ◽  
...  
Keyword(s):  
Glycine Max ◽  
Microrna Targets ◽  
Secondary Sirna

scholarly journals SKI2 mediates degradation of RISC 5′-cleavage fragments and prevents secondary siRNA production from miRNA targets inArabidopsis

2015 ◽  
Vol 43 (22) ◽  
pp. 10975-10988 ◽  
Author(s):  
Anja Branscheid ◽  
Antonin Marchais ◽  
Gregory Schott ◽  
Heike Lange ◽  
Dominique Gagliardi ◽  
...  
Keyword(s):  
Mirna Targets ◽  
Secondary Sirna

Secondary siRNAs Result from Unprimed RNA Synthesis and Form a Distinct Class

Science ◽  
2006 ◽  
Vol 315 (5809) ◽  
pp. 244-247 ◽  
Author(s):  
Titia Sijen ◽  
Florian A. Steiner ◽  
Karen L. Thijssen ◽  
Ronald H. A. Plasterk
Keyword(s):  
Rna Interference ◽  
Small Rnas ◽  
Rna Synthesis ◽  
Argonaute Protein ◽  
Single Nucleotide ◽  
Distinct Class ◽  
C Elegans ◽  
Secondary Sirnas ◽  
Secondary Sirna ◽  
Rnai Response

In Caenorhabditis elegans, an effective RNA interference (RNAi) response requires the production of secondary short interfering RNAs (siRNAs) by RNA-directed RNA polymerases (RdRPs). We cloned secondary siRNAs from transgenic C. elegans lines expressing a single 22-nucleotide primary siRNA. Several secondary siRNAs start a few nucleotides downstream of the primary siRNA, indicating that non–RISC (RNA-induced silencing complex)–cleaved mRNAs are substrates for secondary siRNA production. In lines expressing primary siRNAs with single-nucleotide mismatches, secondary siRNAs do not carry the mismatch but contain the nucleotide complementary to the mRNA. We infer that RdRPs perform unprimed RNA synthesis. Secondary siRNAs are only of antisense polarity, carry 5′ di- or triphosphates, and are only in the minority associated with RDE-1, the RNAi-specific Argonaute protein. Therefore, secondary siRNAs represent a distinct class of small RNAs. Their biogenesis depends on RdRPs, and we propose that each secondary siRNA is an individual RdRP product.

scholarly journals 22-nucleotide RNAs trigger secondary siRNA biogenesis in plants

2010 ◽  
Vol 107 (34) ◽  
pp. 15269-15274 ◽  
Author(s):  
H.-M. Chen ◽  
L.-T. Chen ◽  
K. Patel ◽  
Y.-H. Li ◽  
D. C. Baulcombe ◽  
...  
Keyword(s):  
Secondary Sirna

scholarly journals Genome-wide CRISPR screening identifies new regulators of glycoprotein secretion

2020 ◽  
Vol 4 ◽  
pp. 119
Author(s):  
Stephanie Popa ◽  
Julien Villeneuve ◽  
Sarah Stewart ◽  
Esther Perez Garcia ◽  
Anna Petrunkina Harrison ◽  
...  
Keyword(s):  
Protein Secretion ◽  
Subcellular Localisation ◽  
Novel Genes ◽  
Genome Wide ◽  
Fundamental Process ◽  
Crispr Screen ◽  
Secondary Sirna ◽  
Golgi Structure ◽  
Glycoprotein Secretion ◽  
Selection Of

Background: The fundamental process of protein secretion from eukaryotic cells has been well described for many years, yet gaps in our understanding of how this process is regulated remain. Methods: With the aim of identifying novel genes involved in the secretion of glycoproteins, we used a screening pipeline consisting of a pooled genome-wide CRISPR screen, followed by secondary siRNA screening of the hits to identify and validate several novel regulators of protein secretion. Results: We present approximately 50 novel genes not previously associated with protein secretion, many of which also had an effect on the structure of the Golgi apparatus. We further studied a small selection of hits to investigate their subcellular localisation. One of these, GPR161, is a novel Golgi-resident protein that we propose maintains Golgi structure via an interaction with golgin A5. Conclusions: This study has identified new factors for protein secretion involved in Golgi homeostasis.

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