RIP-Assay Kit for microRNA

  • Target RNP Complex
  • Code # RN1005
  • Size 10 Assays
  • Price
    $351.49
Specifications

Background

Discovery of RNA interference (RNAi) has given a great boost to research on functional RNA and post-transcriptional regulation. RNAi, which plays a central role in sequence-specific gene silencing in eukaryotic cells, depends on the functions of RNA-induced silencing complex (RISC) composed of small non-coding RNAs (ncRNAs) and proteins. The main classes of small ncRNA are short interfering RNAs (siRNAs), microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs). siRNAs are generated by cleavage of exogenous long double-stranded RNA precursors in response to viral infection or artificial introduction. In contrast, miRNAs are generated from endogenous transcripts containing stem-loop structures. The siRNAs and miRNAs processed by Dicer, which functions as a ribonuclease III enzyme, are incorporated into the RISC in order to silence the specific mRNAs based on partial sequence complementarity between the small RNAs and the 3’ untranslated regions (UTRs) of the mRNAs. Argonaute family proteins are a core component of RISC and divided into the AGO and PIWI subfamilies. siRNAs and miRNAs are loaded onto AGO proteins whereas piRNAs are loaded onto PIWI proteins. Each member of the family proteins functions as a silencer to inactivate their target mRNAs. Hundreds of miRNA species have been discovered in animals and plants, many of which exhibit temporally and spatially controlled expression. One approach to investigate the biological functions of miRNAs has been to identify their targets. The miRNA target predictions are based on computational analyses of complementary sequence elements that are refined by considering evolutionary homologies across multiple species. While a variety of computational approaches and algorithms have been used to make such predictions, it is not certain that each miRNA gains functional access to these targeted mRNAs in the cell under a given set of conditions. RIP-Chip (ribonucleoprotein immunoprecipitation-microarray profiling) is a biochemical approach to identify the composition and organization of endogenous mRNAs, miRNAs and RNA binding proteins (RBPs) within messenger ribonucleoprotein (mRNP) complexes. RIP-Chip has been successfully employed to isolate AGO-containing RNPs by immunopurification with anti-AGO antibodies. When the co-isolated miRNA and mRNA subpopulations are analyzed using the computational predictions of conserved seed sequences, this approach provides a powerful tool to identify functional miRNA targets based on their physical interaction in vivo. RBPs have been reported to bind to mRNAs that encode functionally related proteins, and coordinately regulate these mRNAs during cellular processes. The RIP-Chip approach can isolate functionally related mRNAs. Since miRNAs can be co-immunoprecipitated with those mRNAs, the RIP-Chip approach can also isolate miRNAs that regulate specific group of mRNAs that are functionally related.
  • Components:
    • mi-Lysis Buffer
    • mi-Wash Buffer
    • Normal Rabbit IgG
    • High-Salt Solution
    • mi-Solution I
    • mi-Solution II
    • mi-Solution III
    • mi-Solution IV
    • Gel Extraction Buffer
    • 3M NaOAc
    • miSPIKETM 

     

  • Description:

    RIP-Assay Kit for microRNA is optimized to immunochemically isolate endogenous miRNAs, mRNAs and RBPs within mRNP complexes. The kit is designed to isolate cellular miRNAs that being incorporated into the RISC and/or to isolate unique group of miRNAs that bind to specific group of mRNAs encoding functionally related proteins.

  • Product Type:
    Kit
  • Short Description:

    RIP-Assay Kit.

  • Size:
    10 Assays
  • Storage Temperature:
    4°C
  • Target:
    RNP Complex
Citations
  1. Comincini S et al. microRNA-17 regulates the expression of ATG7 and modulates the autophagy process, improving the sensitivity to temozolomide and low-dose ionizing radiation treatments in human glioblastoma cells. Cancer Biol Ther. 14, 574-86 (2013),
  2. Degrauwe N et al. The RNA Binding Protein IMP2 Preserves Glioblastoma Stem Cells by Preventing let-7 Target Gene Silencing. Cell Rep. 15, 1634-47 (2016),
  3. Fu A et al. Targetome profiling and functional genetics implicate miR-618 in lymphomagenesis. Epigenetics 9, 730-7 (2014),
  4. He T et al. MicroRNA-542-3p inhibits tumour angiogenesis by targeting angiopoietin-2. J Pathol. 232, 499-508 (2014),
  5. Hoffman AE et al. Targetome profiling, pathway analysis and genetic association study implicate miR-202 in lymphomagenesis. Cancer Epidemiol Biomarkers Prev. 22, 327-36 (2013),
  6. Madhyastha R et al. NFkappaB activation is essential for miR-21 induction by TGFβ1 in high glucose conditions. Biochem Biophys Res commun. 451, 615-21 (2014)
     ,
  7. Marchesi N et al. Autophagy is modulated in human neuroblastoma cells through direct exposition to low frequency electromagnetic fields. J Cell Physiol. 229, 1776-86 (2014),
  8. Uchino K et al. Therapeutic Effects of MicroRNA-582-5p and -3p on the Inhibition of Bladder Cancer Progression. Mol Ther. 21, 610-9 (2013)
  9. The RNA Binding Protein IMP2 Preserves Glioblastoma Stem Cells by Preventing let-7 Target Gene Silencing. Derauwe N et al. Cell Rep. (2016)