Product CartmicroRNA Processing Factors Expression Vectors
miR-Snares™: Small Non-coding RNA Enrichment Systems
Co-immunopurify microRNA processing factors DGCR8, DICER, AGO family to enrich for microRNAs, targeted mRNAs, and interacting proteins
• Lentivector-based expression of human and mouse microRNA processing factors
• Overexpressed protein factors contain N-terminal HA tag for immunopurification (IP pull-downs)
• Choose from either RFP or Puro markers
• All expression constructs fully sequence-verified and protein expression validated
• Clone and discover low abundance microRNAs
• Profile microRNAs active in RISC complexes
• Uncover novel RISC-associated proteins
• Enrich for RISC-associated mRNAs for microRNA target identification
How to use miR-Snares
1. Transfect or transduce the miR-Snare DNA
2. Immunopurify the protein factor
3. Enrich and discover new small RNAs, or identify mRNAs driven to RISC complexes, or study RISC-associated proteins
miR-Snare Expression Lentivector Information
The miR-Snare constructs are lentiviral vectors containing the microRNA processing factor gene cloned as a translational fusion with an N-terminal hemagglutinin epitope YPYDVPDYA (HA tag). Protein expression is driven by the robust EF1alpha promoter. The construct also contains a downstream T2A peptide ribosomal shift sequence enabling the co-expression of either RFP (fluorescent marker) or the Puro gene for puromycin resistance selection. All constructs are fully sequenced verified for accurate coding content.
Validation of miR-Snare Constructs
All miR-Snare constructs are expression validated. Expression of the specific miR-Snare factor was confirmed through transfection into HEK293 cells with Western blot analysis of protein lysates performed after 48 hours. The protein bands were detected using an anti-HA primary antibody.
The miR-Snare constructs bearing the co-expressed RFP gene were tested using transfection into HEK293 cells and visualized using fluorescent microscopy after 48 hours. Clear RFP signals could be visualized and the expression constructs were non-toxic.
References
Hendrickson DG et al. (2008) Systematic Identification of mRNAs Recruited to Argonaute 2 by Specific microRNAs and Corresponding Changes in Transcript Abundance. PLoS ONE 3(5): e2126.
Fedor V. Karginov et al. (2007) A biochemical approach to identifying microRNA targets. Proc Natl Acad Sci U S A. 104(49): 19291–19296.
Meister G. et al. (2005) Identification of Novel Argonaute-Associated Proteins. Current Biology 15(23):2149 - 2155.
Maniataki E and Mourelatos Z (2005) Human mitochondrial tRNAMet is exported to the cytoplasm and associates with the Argonaute 2 protein. RNA 11(6): 849–852.
Savas JN et al. (2008) Huntington´s disease protein contributes to RNA-mediated gene silencing through association with Argonaute and P bodies. Proc Natl Acad Sci U S A. 105(31):10820-5.
Shipping Information
All constructs are delivered as endotoxin free plasmid DNA.
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| Description | Cat# | Size | Price | ||
|---|---|---|---|---|---|
| pHA-hAGO1-Puro Human AGO1 Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA704A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hAGO1-RFP Human AGO1 Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA704B-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hAGO2-Puro Human AGO2 Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA703A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hAGO2-RFP Human AGO2 Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA703B-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hAGO4-Puro Human AGO4 Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA705A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hAGO4-RFP Human AGO4 Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA705B-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hDGCR8-Puro Human DGCR8 Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA701A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hDGCR8-RFP Human DGCR8 Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA701B-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hDICER-Puro Human Dicer Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA702A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-hDICER-RFP Human Dicer Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA702B-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-mAGO1-Puro Mouse AGO1 Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA801A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-mAGO1-RFP Mouse AGO1 Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA801B-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-mAGO2-Puro Mouse AGO2 Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA802A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-mAGO2-RFP Mouse AGO2 Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA802B-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-mAGO3-Puro Mouse AGO3 Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA803A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-mAGO3-RFP Mouse AGO3 Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA803B-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-mAGO4-Puro Mouse AGO4 Expression Vector (HA-tag for Immunopurification, Puromycin Resistance Marker) | RA804A-1-SBI | 10 ug | 732 € | DETAILS |
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| pHA-mAGO4-RFP Mouse AGO4 Expression Vector (HA-tag for Immunopurification, RFP Marker for Cell Sorting) | RA804B-1-SBI | 10 ug | 732 € | DETAILS |
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