GeneArt™ CRISPR Nuclease Vector with CD4 Enrichment Kit
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GeneArt™ CRISPR Nuclease Vector with CD4 Enrichment Kit
Invitrogen™

GeneArt™ CRISPR Nuclease Vector with CD4 Enrichment Kit

The GeneArt™ CRISPR Nuclease Vector with CD4 Enrichment Kit is a vector system for expression of the functional components neededRead more
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Catalog number A21175
Price (USD)
806.00
Each
Add to cart
Price (USD)
806.00
Each
Add to cart
The GeneArt™ CRISPR Nuclease Vector with CD4 Enrichment Kit is a vector system for expression of the functional components needed for CRISPR/Cas9 genome editing in mammalian cells with a CD4 reporter. The CD4 reporter enables bead-based enrichment, an option for magnetic bead-based sorting/enrichment of Cas9 & CRISPR expressing cells using Dynabeads™ CD4 magnetic beads. Transfection efficiency can also be tracked using anti-CD4 fluorescent antibodies. The linearized GeneArt™ CRISPR nuclease vectors provide a rapid and efficient way to clone double-stranded oligonucleotides encoding a crRNA representing a desired target into an expression cassette that allows sequence-specific targeting of the Cas9 nuclease. A version with competent cells is also available (Cat. No. A21177).

The GeneArt™ CRISPR Nuclease Vector system offers:

• Easy–to-design genome engineering system
• Affordable, ready-to-use cloning vectors
• Enrichment for hard to transfect or difficult to modify cell lines

All-in-one Vector System for CRISPR-based Genome Editing
The GeneArt™ CRISPR Nuclease Vector kit offers a simple, ready-to-use, all-in-one expression vector system consisting of both a Cas9 nuclease expression cassette and a guide RNA (gRNA) cloning cassette for rapid and efficient cloning of DNA that encodes target-specific CRISPR RNA (crRNA). This system allows you to edit and engineer a genomic locus of choice in a sequence-specific manner from a single plasmid. After relevant targets have been identified with fast and easy-to-use GeneArt™ CRISPR vectors, the biologically relevant mutations can be precisely created with GeneArt™ Precision TALs, with high specificity and low off-target effects.

Need assistance with CRISPR gRNA design?
Our new CRISPR Search & Design tool allows you to search our database of >600,000 predesigned CRISPR gRNAs in human and mouse genes or analyze your sequence of interest for de novo gRNA designs using our proprietary algorithms. CRISPR sequences are optimized for gene knockout and target the first three transcribed exons for each gene. Search results include the top 6 CRISPR sequences with PAM sites, exon maps with gRNA binding sites, and potential off-target binding sites for each CRISPR sequence. The tool will design the correct gRNA format for your preferred CRISPR-Cas9 product, including oligos for your GeneArt™ CRISPR Nuclease Vector. Start designing today >
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Cloning MethodRestriction Enzyme/MCS
Detection MethodPrimer-probe
FormLiquid
FormatKit
Reaction SpeedFast
TechniqueCRISPR-Cas9
Product LineGeneArt™
Product TypeCrispr Nuclease Vector With CD4 Enrichment Kit
PromoterU6, CMV
Protein TagCD4 fusion
Reporter GeneCD4
Sufficient For10 Reactions
For Use With (Application)Gene Expression, Cloning
No. of Reactions10 Reactions
Overhang3' overhangs
Starting MaterialOligos (DNA)
Unit SizeEach
Contents & Storage
Contains:
• CRISPR CD4 Nuclease Vector, linearized
• 10X Oligonucleotide Annealing Buffer
• DNase/RNase-free Water
• 5X Ligation Buffer
• T4 DNA Ligase
• U6 Forward Sequencing Primer
• Control Double-stranded Oligonucleotide

Store in freezer (-5 to -30°C).

Frequently asked questions (FAQs)

What is CRISPR-STOP?

CRISPR-STOP is a method of inserting STOP codon sequences to generate knockouts.

Please refer to the following article: CRISPR-STOP: gene silencing through base-editing-induced nonsense mutations.

Find additional tips, troubleshooting help, and resources within our Genome Editing Support Center.

How do I check for off-target effects in my CRISPR-modified cell lines?

The only complete way to confirm that there are no off-target effects is to sequence the entire genome of your cell. Alternatively, a less thorough means of checking for off-target editing is to perform targeted sequencing of sequences with the highest probability of off-target effects (i.e., most similar to your CRISPR target region).

How many guide RNAs do you recommend designing against my desired edit locus?

A single guide RNA (gRNA) is all that is required for targeting, but we do recommend testing 2-3 gRNAs against each locus being targeted for cleavage. Testing multiple gRNAs increases the chances of finding a gRNA with high editing efficiency, which will reduce the screening time required to identify the clone of interest.

What transfection methods do you recommend when working with your CRISPR products?

For transfecting of the Cas9 protein, we would recommend using the Neon transfection system or Lipofectamine CRISPRMAX Cas9 Transfection Reagent. For transfection of mRNA, we would recommend using Lipofectamine MessengerMAX Transfection Reagent. For transfection of CRISPR vectors, we would recommend using Lipofectamine 3000 Transfection reagent.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Do you have CRISPR products optimized for plants?

Our CRISPR products are optimized for mammalian systems, and have not been optimized for plant systems. However, we know researchers are doing that kind of work. While we have not tested our CRISPR products for plants, our new protein format would be ideal since it does not need to be translated and transcribed in the cell, so no plant-specific promoters required.