Tie-1 CRISPR Plasmids (m)

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- Target species: mouse; for corresponding human product, see Tie-1 CRISPR Plasmids (h)
- CRISPR/Cas9 KO Plasmids consists of Tie-1-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
- 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

Species	Gene Name	Gene ID	Chromosome Location	Isoform (mRNA) Accession Number	Protein Accession Number	OMIM™ Number
Mouse	Tie1	Entrez Gene ID: 21846	4 D2.1	NM_011587	UniProt ID: Q06806	N/A

Description

20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
Tie-1 CRISPR/Cas9 Knockout (KO) Plasmid (m) consists of a pool of three plasmids each encoding the Cas9 nuclease and a Tie-1-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 Tie-1 HDR Plasmid (m) (sc-423394-HDR) is recommended
contact Technical Service for availability of control antibody

See Protocol Online.

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Description

20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
Tie-1 HDR Plasmid (m) is recommended for co-transfection with Tie-1 CRISPR/Cas9 KO Plasmid (m)
Tie-1 HDR Plasmid (m) 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 Tie-1 CRISPR/Cas9 KO Plasmid (m)
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
Tie-1 Double Nickase Plasmid (m) 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 Tie-1 Double Nickase Plasmid (m2) differ from the paired gRNAs in Tie-1 Double Nickase Plasmid (m)

See Protocol Online.

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Description

20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
Tie-1 CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
Tie-1 CRISPR Activation Plasmid (m) 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

See Protocol Online.

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Description

200 µl of transduction-ready, high-titer CRISPR/dCas9 Lentiviral Activation Particles
Tie-1 Lentiviral Activation Particles (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically and efficiently upregulate gene expression via lentiviral transduction of cells
Tie-1 Lentiviral Activation Particles (m) 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 Tie-1 Lentiviral Activation Particles (m2) encodes a target-specific 20 nt guide RNA that differs from the guide RNA in Tie-1 Lentiviral Activation Particles (m)

See Protocol Online.

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