miR and PIWI-SNaREs™
Immunopurify microRNA and piRNA complexes
and their associated RNAs
HA-tagged microRNA and piRNA factors
- DGCR8 (Pasha)
- DICER
- Argonautes 1, 2, 3, 4
- Human PIWI factors
Features and Benefits
- Lentivector-based cDNA expression of piRNA-binding (PIWI) 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
- The expression constructs are protein expression validated
- Discover new piRNAs
- Identify new epigenetic RNAs and associated protein factors
- Enrich for active PIWI complex-associated RNAs
Lentivector-based PIWI protein expression
The PIWI-Snare constructs contain the PIWI factor genes 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.
All PIWI-Snare constructs are expression validated
Expression of the specific PIWI-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.
Co-expression of RFP fluorescent marker validation
The PIWI-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.
