Branch-PCR Constructed Stable shRNA Transcription Nanoparticles Have Long-Lasting RNAi Effect

ChemBioChem ◽  
2016 ◽  
Vol 17 (11) ◽  
pp. 1038-1042 ◽  
Author(s):  
Jianbing Liu ◽  
Runyu Wang ◽  
Dejun Ma ◽  
Yanyan Li ◽  
Chao Wei ◽  
...  
Keyword(s):  
Rnai Effect

Utilization of a PNA-peptide conjugate to induce a cancer protease-responsive RNAi effect

RSC Advances ◽  
2015 ◽  
Vol 5 (104) ◽  
pp. 85816-85821
Author(s):  
Eun Kyung Lee ◽  
Chan Woo Kim ◽  
Hiroyuki Kawanami ◽  
Akihiro Kishimura ◽  
Takuro Niidome ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Cancer Cell ◽  
Peptide Nucleic Acid ◽  
Specific Protease ◽  
Rnai Effect

We designed a new siRNA system which turns on RNAi responding to a cancer cell-specific protease by using a peptide nucleic acid (PNA)-peptide conjugate.

scholarly journals Construction of Henosepilachna Vigintioctomaculata cDNA Library and Identification of dsRNA Injection–Induced RNAi Effect

2020 ◽  
Vol 65 (1) ◽  
pp. 79-95
Author(s):  
Jeong Hee KIM ◽  
Hye Ri KWON ◽  
Yong Man YU ◽  
Chisa YASUNAGA–AOKI ◽  
Young Nam YOUN
Keyword(s):  
Cdna Library ◽  
Dsrna Injection ◽  
Rnai Effect

scholarly journals A study on the fundamental factors determining the efficacy of siRNAs with high C/G contents

2008 ◽  
Vol 13 (2) ◽  
Author(s):  
Jie-Ying Liao ◽  
James Yin ◽  
Fang Chen ◽  
Tie-Gang Liu ◽  
Jia-Chang Yue
Keyword(s):  
Gene Silencing ◽  
Structure And Function ◽  
Guide Strand ◽  
Target Mrna ◽  
Target Mrnas ◽  
Passenger Strand ◽  
Target Sequences ◽  
And Function ◽  
Necessary And Sufficient ◽  
Rnai Effect

AbstractAlthough there are many reports about the efficacy of siRNAs, it is not clear whether those siRNAs with high C/G contents can be used to silence their target mRNAs efficiently. In this study, we investigated the structure and function of a group of siRNAs with high C/G contents. The results showed that single siRNAs against the Calpain, Otoferlin and Her2 mRNAs could induce different silencing effects on their targets, suggesting that the accessibility to target sequences influences the efficacy of siRNA. Unexpectedly, a single siRNA could target its cognate sequence in the 3’UTR of EEF1D or the 5’UTR of hTRF2 or CDC6. Their interaction induced different modes of gene silencing. Furthermore, the introduction of mutations into the 3’ end of the passenger strand showed that the position and number of mutated nucleotides could exert some influence on the efficacy of siRNA. However, these mutations did not completely block the passenger strand from exerting its RNAi effect. Interestingly, our findings also indicated that the target mRNA might play essential roles in maintaining or discarding the guide strand in RISCs. Thus, the conclusion could be drawn that favorable siRNA sequences, accessible target structures and the fast cleavage mode are necessary and sufficient prerequisites for efficient RNAi.

scholarly journals RNA interference of an orthologue of Dicer of Meloidogyne incognita alludes to the gene’s importance in nematode development

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sadia Iqbal ◽  
Michael G. K. Jones ◽  
John Fosu-Nyarko
Keyword(s):  
Rna Interference ◽  
Small Rna ◽  
Meloidogyne Incognita ◽  
Rna Processing ◽  
Essential Role ◽  
Juvenile Stage ◽  
Coding Sequences ◽  
Adult Females ◽  
Rnai Effect ◽  
Nematode Development

AbstractDicers and dicer-like enzymes play an essential role in small RNA processing in eukaryotes. Nematodes are thought to encode one dicer, DCR-1; only that for Caenorhabditis spp. is well-characterised. Using genomic sequences of eight root-knot nematodes (Meloidogyne spp.), we identified putative coding sequences typical of eukaryotic DICERS. We noted that the primary and secondary structures of DICERS they encode were different for different Meloidogyne species and even for isolates of the same species, suggesting paralogy for the gene. One of the genes for M. incognita (Midcr-1.1) expressed in eggs, juvenile stage 2 and adults, with the highest expression in the adult females. All the Meloidogyne DICERS had seven major domains typical of those for Caenorhabditis spp. and humans with very similar protein folding. RNAi of Midcr-1.1 in J2s using seven dsRNAs, each based on sequences encoding the domains, induced mild paralysis but measurable knockdown was detected in J2s treated with five of the dsRNAs. For four of the dsRNAs, the RNAi effect lasted and reduced the nematode’s infectivity. Also, host plant delivery of dsRNAs complementary to coding sequences of the Dicer Dimerisation domain impaired development, reducing nematode infection by 71%. These results confirm the importance of the gene to nematode health.

scholarly journals Length-dependent accumulation of double-stranded RNAs in plastids affects RNA interference efficiency in the Colorado potato beetle

2020 ◽  
Vol 71 (9) ◽  
pp. 2670-2677 ◽  
Author(s):  
Wanwan He ◽  
Wenbo Xu ◽  
Letian Xu ◽  
Kaiyun Fu ◽  
Wenchao Guo ◽  
...  
Keyword(s):  
Rna Interference ◽  
Colorado Potato Beetle ◽  
Plant Protection ◽  
Larval Growth ◽  
High Accumulation ◽  
Potato Plants ◽  
Potato Beetle ◽  
Rnai Response ◽  
Rnai Effect

Abstract Transplastomic potato plants expressing double-stranded RNA (dsRNA) targeted against essential genes of the Colorado potato beetle (CPB) can be lethal to larvae by triggering an RNA interference (RNAi) response. High accumulation levels of dsRNAs in plastids are crucial to confer an efficient RNAi response in the insects. However, whether length and sequence of the dsRNA determine the efficacy of RNAi and/or influence the level of dsRNA accumulation in plastids is not known. We compared the RNAi efficacy of different lengths of dsRNA targeted against the CPB β-Actin gene (ACT) by feeding in vitro-synthesized dsRNAs to larvae. We showed that, while the 60 bp dsRNA induced only a relatively low RNAi response in CPB, dsRNAs of 200 bp and longer caused high mortality and similar larval growth retardation. When the dsRNAs were expressed from the plastid (chloroplast) genome of potato plants, we found that their accumulation were negatively correlated with length. The level of dsRNA accumulation was positively associated with the observed mortality, suppression of larval growth, and suppression of target gene expression. Importantly, transplastomic potato plants expressing the 200 bp dsRNA were better protected from CPB than plants expressing the 297 bp dsRNA, the best-performing line in our previous study. Our results suggest that the length of dsRNAs is an important factor that influences their accumulation in plastids and thus determines the strength of the insecticidal RNAi effect. Our findings will aid the design of optimized dsRNA expression constructs for plant protection by plastid-mediated RNAi.

Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system

2000 ◽  
Vol 113 (18) ◽  
pp. 3321-3328 ◽  
Author(s):  
P. Bastin ◽  
K. Ellis ◽  
L. Kohl ◽  
K. Gull
Keyword(s):  
Rna Interference ◽  
Trypanosoma Brucei ◽  
Construction Process ◽  
Cellular Processes ◽  
Cytoskeletal Structure ◽  
Interference System ◽  
Turn Over ◽  
Fast Acting ◽  
Inducible Rnai ◽  
Rnai Effect

The African trypanosome, Trypanosoma brucei possesses a large and unique intraflagellar structure called the paraflagellar rod (PFR). The PFR is composed of 2 major proteins, PFRA and PFRC. We have generated an inducible mutant trypanosome cell line (snl-2) that expresses linked inverted copies of a PFRA gene, capable of forming a PFRA double-stranded (ds) RNA. When expression of this dsRNA was induced, new PFRA RNA and PFRA protein quickly disappeared and PFR construction was affected, resulting in cell paralysis. This inducible RNA interference (RNAi) effect was fast-acting, heritable and reversible. It allowed us to demonstrate that PFR proteins are able to enter both mature and growing flagella but appear to concentrate differentially in new flagella because of the construction process. The PFR is constructed by a polar assembly process at the distal end of the flagellum resulting in a stable cytoskeletal structure with low turn-over. The inducible RNAi approach will have widespread applicability in studies of gene function and cellular processes in parasites.

scholarly journals Length-dependent accumulation of double-stranded RNAs in plastids affects RNA interference efficiency in the Colorado potato beetle

2019 ◽  
Author(s):  
Wanwan He ◽  
Wenbo Xu ◽  
Letian Xu ◽  
Kaiyun Fu ◽  
Wenchao Guo ◽  
...  
Keyword(s):  
Rna Interference ◽  
Colorado Potato Beetle ◽  
Plant Protection ◽  
Larval Growth ◽  
High Accumulation ◽  
Potato Plants ◽  
Potato Beetle ◽  
Rnai Response ◽  
Rnai Effect

AbstractTransplastomic potato plants expressing double-stranded RNA (dsRNA) targeted against essential genes of the Colorado potato beetle (CPB) can be lethal to larvae by triggering an RNA interference (RNAi) response. High accumulation levels of dsRNAs in plastids are crucial to confer an efficient RNAi response in the insects. However, whether length and sequence of the dsRNA determine the efficacy of RNAi and/or influence the level of dsRNA accumulation in plastids is not known. Here we compared the RNAi efficacy of different lengths of dsRNA targeted against the CPB β–Actin gene (ACT) by feeding in vitro-synthesized dsRNAs to larvae. We show that, while the 60 bp dsRNA induced only a relatively low RNAi response in CPB, dsRNAs of 200 bp and longer caused high mortality and similar larval growth retardation. When the dsRNAs were expressed from the plastid (chloroplast) genome of potato plants, we found that their accumulation levels were correlated with length. dsRNA accumulation levels were positively associated with the observed mortality, suppression of larval growth and suppression of target gene expression. Importantly, transplastomic potato plants expressing the 200 bp dsRNA were better protected from CPB than plants expressing the 297 bp dsRNA, the best-performing line in our previous study. Our results suggest that the length of dsRNAs is an important factor that influences their accumulation levels in plastids and thus determines the strength of the insecticidal RNAi effect. Our findings will aid the design of optimized dsRNA expression constructs for plant protection by plastid-mediated RNAi.

scholarly journals Intracellular stability of 2′-OMe-4′-thioribonucleoside modified siRNA leads to long-term RNAi effect

2012 ◽  
Vol 40 (12) ◽  
pp. 5787-5793 ◽  
Author(s):  
Mayumi Takahashi ◽  
Chisato Nagai ◽  
Hiroto Hatakeyama ◽  
Noriaki Minakawa ◽  
Hideyoshi Harashima ◽  
...  
Keyword(s):  
Rnai Effect
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