Each CRISPR/Cas9 Knockout (KO) Plasmid product consists of a pool of 3 plasmids designed to ensure identification and cleavage of a specific gene for maximum knockout efficiency. Each plasmid encodes a unique 20 nt sequence derived from the GeCKO (v2) library.
Topo IIα CRISPR Plasmids (h)
- Target species: human; for corresponding mouse product, see Topo IIα CRISPR Plasmids (m)
- CRISPR/Cas9 KO Plasmids consists of Topo IIα-specific 20 nt guide RNA sequences derived from the GeCKO (v2) library
- For CRISPR gene knockout, gRNA sequences direct the Cas9 protein to induce a site-specific double strand break (DSB) in the genomic DNA
- Target-specific CRISPR Plasmids for both gene knockout and activation are available. Please refer to the detailed product information in the tabs below
- Gene knockdown or activation can be assayed using Topo IIα Antibody (G-6): sc-166934
- All products are provided as transfection-ready, purified plasmid DNA, except the Lentiviral Activation Particles which have been prepackaged as Lentiviral Particles for use with hard-to-transfect cells
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Ordering Information
CRISPR/Cas9 KO Plasmid
HDR Plasmid
Double Nickase Plasmids
CRISPR Activation Plasmids
Lentiviral Activation Particles
Recommended CRISPR and HDR Support Reagents
Recommended Control Products
SEE ALSO...
Topo II α Antibodies for analysis of cellular responses to Topo II α CRISPR Products
Learn more about the CRISPR System
CRISPR/Cas9 KO plasmids, Double Nickase Plasmids and CRISPR/dCas9 Activation plasmids are considered a "Licensed Product" and are to be used in accordance with the Limited/Label License. Click here for details
Description
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIα CRISPR/Cas9 Knockout (KO) Plasmid (h) consists of a pool of three plasmids each encoding the Cas9 nuclease and a Topo IIα-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency
- gRNA sequences are derived from the GeCKO (v2) library and direct the Cas9 protein to induce a site-specific double strand break (DSB) in the genomic DNA
- For selection of cells containing a successful Cas9-induced DSB, co-transfection with Topo IIα HDR Plasmid (h) (sc-400869-HDR) is recommended
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Topo IIα Antibody (G-6): sc-166934
See Protocol Online.
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Description
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIα HDR Plasmid (h) is recommended for co-transfection with Topo IIα CRISPR/Cas9 KO Plasmid (h)
- Topo IIα HDR Plasmid (h) consists of a pool of 2-3 plasmids, each containing a homology-directed DNA repair (HDR) template corresponding to the cut sites generated by the Topo IIα CRISPR/Cas9 KO Plasmid (h)
- Each HDR plasmid contains two 800 bp homology arms designed to specifically bind to the genomic DNA surrounding the corresponding Cas9-induced double strand DNA break site
- Each HDR Plasmid inserts a puromycin resistance gene to enable selection of stable knockout (KO) cells and an RFP (Red Fluorescent Protein) gene to visually verify transfection
- The puromycin resistance and RFP genes are flanked by two LoxP sites to allow for further processing by the Cre Vector sc-418923
See Protocol Online.
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Description
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIα Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- The paired gRNAs encoded by Topo IIα Double Nickase Plasmid (h2) differ from the paired gRNAs in Topo IIα Double Nickase Plasmid (h)
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Topo IIα Antibody (G-6): sc-166934
See Protocol Online.
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Description
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIα CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Topo IIα CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- Following transfection, gene activation efficiency can be assayed by WB, IF or IHC using antibody: Topo IIα Antibody (G-6): sc-166934
See Protocol Online.
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Description
- 200 µl of transduction-ready, high-titer CRISPR/dCas9 Lentiviral Activation Particles
- Topo IIα Lentiviral Activation Particles (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically and efficiently upregulate gene expression via lentiviral transduction of cells
- Topo IIα Lentiviral Activation Particles (h) contain the following SAM Activation elements: a deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, an MS2-p65-HSF1 fusion protein and a target-specific 20 nt. guide RNA. They also contain the blasticidin, hygromycin and puromycin resistance genes
- Upon transduction, the SAM complex binds to a site-specific region approximately 200-250 nt. upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- The guide RNA in the Topo IIα Lentiviral Activation Particles (h2) encodes a target-specific 20 nt guide RNA that differs from the guide RNA in Topo IIα Lentiviral Activation Particles (h)
- Following transfection, gene activation efficiency can be assayed by WB, IF or IHC using antibody: Topo IIα Antibody (G-6): sc-166934
See Protocol Online.
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Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Topo IIα CRISPR/Cas9 KO Plasmid (h) | sc-400869 | 20 µg | $390.00 | |||
Topo IIα HDR Plasmid (h) | sc-400869-HDR | 20 µg | $437.00 | |||
Topo IIα Double Nickase Plasmid (h) | sc-400869-NIC | 20 µg | $402.00 | |||
Topo IIα Double Nickase Plasmid (h2) | sc-400869-NIC-2 | 20 µg | $402.00 | |||
Topo IIα CRISPR Activation Plasmid (h) | sc-400869-ACT | 20 µg | $390.00 | |||
Topo IIα CRISPR Activation Plasmid (h2) | sc-400869-ACT-2 | 20 µg | $390.00 | |||
Topo IIα Lentiviral Activation Particles (h) | sc-400869-LAC | 200 µl | $447.00 | |||
Topo IIα Lentiviral Activation Particles (h2) | sc-400869-LAC-2 | 200 µl | $447.00 |