A rapid method to screen putative mRNA targets of any known microRNA

Huang, Yujing; Qi, Ying; Ruan, Qiang; Ma, Yanping; He, Rong; Ji, Yaohua; Sun, Zhengrong

doi:10.1186/1743-422x-8-8

Cited by 43 publications

(35 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As acquired from previous hybrid PCR method [17], we found one potential binding site for miR-134 in the CDS (coding sequence) region of Pak2 gene (The coding region of a gene, also known as the coding sequence or CDS, is that portion of a gene's RNA, composed of exons, that codes for protein). We designed one pair of primers for cloning the fragment containing the potential binding sites of miR-134 (Primers used for the fragment containing were as following: Sense 5 0 GGACTAGT ATG TCT GAT AAC GGA GAA CTG GAAG 3 0 , Antisense: 5 0 CCCAAGCTT TCT CCA GTA ACA GCA TCAA 3 0 ).…”

Section: Plasmid Generationmentioning

confidence: 99%

“…To reveal the possible mechanism of decreased expression of miR-134 in ovarian cancer chemoresistance, we applied hybrid PCR [17] to find the potential target genes for miR-134, and found Pak2 gene was a potential target of miR-134and the bindingsite was located in the CDS region of Pak2 mRNA. Then we found Pak2 protein level was increased in ovarian cancer drug resistant SKOV3-TR30 cells compared with the parental SKOV3 cells while the mRNA level was similar.…”

Section: Discussionmentioning

confidence: 99%

“…To explore the putative target of miR-134, we applied hybrid PCR [17] method and found Pak2 gene was one of the potential targets of miR-134 with one putative target site in the CDS region and the target sequences were highly conserved among different species ( Fig. 2A).…”

Section: Mir-134 Directly Targets Pak2 Cds Regionmentioning

confidence: 99%

See 2 more Smart Citations

Down‐regulated expression of miR‐134 contributes to paclitaxel resistance in human ovarian cancer cells

et al. 2015

View full text Add to dashboard Cite

a b s t r a c tMiR-134 has been reported to have a role in the development and progression of various cancers. In this study, we found that miR-134 expression was significantly decreased in chemo-resistant serous epithelial ovarian cancer (EOC) patients. Over-expression of miR-134 enhanced the sensitivity of SKOV3-TR30 cells to paclitaxel, and increased paclitaxel-induced apoptosis. Further, Pak2 was identified as a direct target of miR-134, and Pak2-specific siRNA increased cell inhibition rate and promoted paclitaxal-induced apoptosis. By regulating Pak2 expression, miR-134 could mediate Bad phosphorylation at Ser112 and Ser136, which affected cell survival and apoptosis. In conclusion, our findings indicate that repression of miR-134 and consequent up-regulation of Pak2 might contribute to paclitaxel resistance.

show abstract

Section: Plasmid Generationmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Down‐regulated expression of miR‐134 contributes to paclitaxel resistance in human ovarian cancer cells

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Target genes of hcmv-miR-UL148D were screened using the hybrid-PCR method as described in a previous study of ours (27). The hybrid-PCR primer of hcmv-miR-UL148D, A1, was designed according to the reverse sequence of hcmv-miR-UL148D (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Since miRNAs exhibit post-trancriptional regulatory effects by binding to target mRNAs, the target mRNA sequences were screened within mRNA-derived cDNA from HCMV-infected cells using hybrid-PCR, which has been experimentally demonstrated to be efficient for screening target genes of any known miRNAs (27). Two main products of hcmv-mir-UL148D obtained from the HCMV-infected cells by hybrid-PCR were ~300 and 500 bp in length.…”

Section: Putative Target Genes Of Hcmv-mir-ul148d Screened By Hybrid-mentioning

confidence: 99%

Identification of immediate early gene X-1 as a cellular target gene of hcmv-mir-UL148D

Wang

Huang

et al. 2013

International Journal of Molecular Medicine

Self Cite

View full text Add to dashboard Cite

Abstract. Human cytomegalovirus (HCMV) is a herpesvirus that causes congenital diseases and opportunistic infections in immunocompromised individuals. Its functional proteins and microRNAs (miRNAs) facilitate efficient viral propagation by altering host cell behavior. The identification of functional target genes of miRNAs is an important step in the study of HCMV pathogenesis. HCMV encodes at least 14 miRNAs, including hcmv-mir-UL148D, which resides in the HCMV UL/b' region. hcmv-mir-UL148D is the only miRNA encoded by the HCMV UL/b' region; however, its targets and functional effects have not yet been eludidated. In this study, hybrid-PCR screening was used to identify target genes and dual luciferase reporter assay was used to evaluate the binding effect of hcmv-miR-UL148D to the 3' untranslated region (3'UTR) of IEX-1. In addition, western blot analysis was used to detect the expression kinetics of IEX-1 protein and apoptosis assay was used to identify the effects of hcmv-miR-UL148D on cell apoptosis. The hybrid-PCR results showed that only one binding site in the 3'UTR of the cellular gene, human immediate early gene X-1 (IEX-1), was completely complementary to an 11 nucleotide (nt) sequence in the 5' terminus of hcmvmir-UL148D, including the entire seed region. The binding site was demonstrated to be functional by dual luciferase reporter assay with a 47% repression of the relative luciferase activity. In an in vitro system of human embryonic kidney 293 (HEK293) cells, the ectopically expressed hcmv-mir-UL148D exhibited a downregulatory effect on IEX-1 expression, and decreased the cell apoptosis induced by transfected IEX-1. Our data demonstrate that hcmv-mir-UL148D targets the cellular gene, IEX-1, downregulating its expression and thus results in anti-apoptotic effects.

show abstract

Interleukin‐17—A multifaceted cytokine in viral infections

Sahu

Biswas

Prajapati

et al. 2021

Journal Cellular Physiology

View full text Add to dashboard Cite

Viral infections are a major threat to the human population due to the lack of selective therapeutic measures. The morbidity and mortality reported worldwide are very alarming against viral pathogens. The proinflammatory environment is required for viral inhibition by initiating the host immune response. The host immune response fights these pathogens by secreting different cytokines. Interleukin‐17 (IL‐17) a proinflammatory cytokine mainly produced by T helper type 17 cells, plays a vital role in the regulation of host immune response against various pathogens, including viruses. However, dysregulated production of IL‐17 induces chronic inflammation, autoimmune disorders, and may lead to cancer. Recent studies suggest that IL‐17 is not only involved in the antiviral immune response but also promotes virus‐mediated illnesses. In this review, we discuss the protective and pathogenic role of IL‐17 against various viral infections. A detailed understanding of IL‐17 during viral infections could contribute to improve therapeutic measures and enable the development of an efficient and safe IL‐17 based immunotherapy.

show abstract

A rapid method to screen putative mRNA targets of any known microRNA

Cited by 43 publications

References 16 publications

Down‐regulated expression of miR‐134 contributes to paclitaxel resistance in human ovarian cancer cells

Down‐regulated expression of miR‐134 contributes to paclitaxel resistance in human ovarian cancer cells

Identification of immediate early gene X-1 as a cellular target gene of hcmv-mir-UL148D

Interleukin‐17—A multifaceted cytokine in viral infections

Contact Info

Product

Resources

About