cd2bp2 c sirna duplex Search Results


cd2bp2-c sirna duplex  (OriGene)
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OriGene cd2bp2-c sirna duplex
Association of TEG-1 with miRISC proteins is conserved in human tissue culture cells. ( A ) SERBP1/PAI-RBP1 (VIG-1 ortholog) and AGO2 (ALG-1 ortholog) are co-IP'd with <t>CD2BP2</t> (TEG-1 ortholog). GFP or GFP-CD2BP2 was transiently expressed in HeLa cells and proteins were co-IP'd using anti-GFP antibodies. SERBP1/PAI-RBP1 and AGO2 are detected when IP'd with anti-GFP antibodies followed by immunoblotting, while the negative control, actin, was not detected. ( B ) Mature let-7a, miR-24, and miR-26a miRNAs are significantly reduced in HeLa cells treated with CD2BP2 siRNA in comparison with a scrambled siRNA treatment by quantitative real time PCR using TaqMan probes. A two-tailed Student's t -test was used to determine the statistical significance: * P < 0.0003. Error bar = standard deviation. ( C ) Proposed model. Free miRNAs are degraded by exonucleases (XRN-1/XRN-2), while miRISC proteins are subjected to proteasomal degradation. Our model suggests that TEG-1 CD2BP2 interacts with miRISC. TEG-1 CD2BP2 helps stabilize miRISC protein components. In the absence of TEG-1 CD2BP2, miRISC protein components are more susceptible to proteasomal degradation (thick arrow), which in turn results in decreased stability of miRNAs.
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Association of TEG-1 with miRISC proteins is conserved in human tissue culture cells. ( A ) SERBP1/PAI-RBP1 (VIG-1 ortholog) and AGO2 (ALG-1 ortholog) are co-IP'd with CD2BP2 (TEG-1 ortholog). GFP or GFP-CD2BP2 was transiently expressed in HeLa cells and proteins were co-IP'd using anti-GFP antibodies. SERBP1/PAI-RBP1 and AGO2 are detected when IP'd with anti-GFP antibodies followed by immunoblotting, while the negative control, actin, was not detected. ( B ) Mature let-7a, miR-24, and miR-26a miRNAs are significantly reduced in HeLa cells treated with CD2BP2 siRNA in comparison with a scrambled siRNA treatment by quantitative real time PCR using TaqMan probes. A two-tailed Student's t -test was used to determine the statistical significance: * P < 0.0003. Error bar = standard deviation. ( C ) Proposed model. Free miRNAs are degraded by exonucleases (XRN-1/XRN-2), while miRISC proteins are subjected to proteasomal degradation. Our model suggests that TEG-1 CD2BP2 interacts with miRISC. TEG-1 CD2BP2 helps stabilize miRISC protein components. In the absence of TEG-1 CD2BP2, miRISC protein components are more susceptible to proteasomal degradation (thick arrow), which in turn results in decreased stability of miRNAs.

Journal: Nucleic Acids Research

Article Title: TEG-1 CD2BP2 controls miRNA levels by regulating miRISC stability in C. elegans and human cells

doi: 10.1093/nar/gkw836

Figure Lengend Snippet: Association of TEG-1 with miRISC proteins is conserved in human tissue culture cells. ( A ) SERBP1/PAI-RBP1 (VIG-1 ortholog) and AGO2 (ALG-1 ortholog) are co-IP'd with CD2BP2 (TEG-1 ortholog). GFP or GFP-CD2BP2 was transiently expressed in HeLa cells and proteins were co-IP'd using anti-GFP antibodies. SERBP1/PAI-RBP1 and AGO2 are detected when IP'd with anti-GFP antibodies followed by immunoblotting, while the negative control, actin, was not detected. ( B ) Mature let-7a, miR-24, and miR-26a miRNAs are significantly reduced in HeLa cells treated with CD2BP2 siRNA in comparison with a scrambled siRNA treatment by quantitative real time PCR using TaqMan probes. A two-tailed Student's t -test was used to determine the statistical significance: * P < 0.0003. Error bar = standard deviation. ( C ) Proposed model. Free miRNAs are degraded by exonucleases (XRN-1/XRN-2), while miRISC proteins are subjected to proteasomal degradation. Our model suggests that TEG-1 CD2BP2 interacts with miRISC. TEG-1 CD2BP2 helps stabilize miRISC protein components. In the absence of TEG-1 CD2BP2, miRISC protein components are more susceptible to proteasomal degradation (thick arrow), which in turn results in decreased stability of miRNAs.

Article Snippet: To study the effect of CD2BP2 down-regulation by siRNA, 2 × 10 5 HeLa-CCL2 cells were transfected with either a universal scrambled negative control siRNA duplex or CD2BP2-C siRNA duplex (5΄-CUCAGGCAGCGAGGAAAUUGGAGGC-3΄ (ORIGENE)) at 10 nM using Lipofectamin 2000 according to the manufacturer's instructions (Life Technologies).

Techniques: Co-Immunoprecipitation Assay, Western Blot, Negative Control, Real-time Polymerase Chain Reaction, Two Tailed Test, Standard Deviation