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Build your own CRISPR solution
Customize your vector
The powerful CRISPR gene editing technology transforms research at an astonishing rate. Read about a five-step approach for DIY CRISPR workflow, with focus on CRISPR-Cas9 vector design, its construction and delivery into cells, possible ways for mutant genotyping and characterization. We’ve refined and optimized each step to help ensure maximum editing efficiency across a broad spectrum of cell types.
The CRISPR-Cas9 system is a two-component system, consisting of the target-specific CRISPR gRNA and Cas9 nuclease. For genome editing to be successful, both the Cas9 and gRNA need to be expressed together in the target cells. We offer the necessary tools for the design and engineering of target-specific gRNA and Cas9 nuclease expression plasmids enabling you to pursue different experimental strategies.
Products for CRISPR-Cas9 design
For designing your vector you may choose between two separate plasmids for Cas9 and gRNA, respectively, or a single plasmid for both genes:
- Plasmid for Cas nuclease expression obtained from open-access resources (e.g., Addgene, the nonprofit global plasmid repository)
- gRNA cloning vector obtained from open-access resources (e.g., Addgene)
- GeneArt CRISPR Nuclease vector for cloning Cas9 and gRNA together
All-in-one plasmid system
Transfect, enrich, screen, and publish by using our GeneArt CRISPR Nuclease Vector Kit. The CRISPR Nuclease system offers a ready-to-use, all-in-one expression vector system with a Cas9 nuclease expression cassette and a guide RNA cloning cassette for fast cloning of a target-specific crRNA. This system allows you to edit and engineer the genomic locus of your choice in a sequence-specific manner from a single plasmid. After relevant targets have been identified with GeneArt CRISPRs, the biologically-relevant mutations can be validated with GeneArt TALs to reduce potential off-targeting.
GeneArt CRISPR Nuclease Vector Kits
GeneArt CRISPR Nuclease Vector Kits are reporter vector systems for expression of the functional components needed for CRISPR-Cas genome editing. The kits make it easy to express noncoding guide RNA (including crRNA and tracrRNA), using a plasmid vector that also expresses Cas9 endonuclease.
The GeneArt CRISPR Nuclease Vector with OFP (orange fluorescent protein) for flow cytometry-based sorting of crRNA-expressing cell populations, whereas GeneArt CRISPR Nuclease Vector with CD4 enables bead-based enrichment of crRNA-expressing cells.
GeneArt CRISPR Nuclease Vectors. (A) GeneArt CRISPR Nuclease: OFP Reporter Plasmid map and features of GeneArt CRISPR Nuclease: OFP Reporter. The vector is supplied linearized between nucleotides 6,732 and 6,752, with 5 bp 5´ overhangs on each strand as indicated. (B) GeneArt CRISPR Nuclease: CD4 Enrichment Plasmid map and features of GeneArt CRISPR Nuclease: CD4 Enrichment. The vector is supplied linearized between nucleotides 7,336 and 7,355, with 5 bp 5´ overhangs on each strand as indicated. 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.
| For constructing your own plasmid for gRNA expression you may need: | |
|---|---|
.jpg) | |
| For easy cloning of novel CRISPR nuclease sequences or subcloning work you may need: | |
.jpg) |
|
| Custom CRISPR for every gene; we'll design your target and clone it for you. Ready-to-transfect. | Contact us |
Related products
Invitrogen TrueTag Donor DNA Kits can help you obtain up to 100% knocked-in cells using our predesigned and validated donor DNA templates.
Looking for our premium genome editing solutions?
Learn more about our first-class genome engineering products, including TrueCut Cas9 v2 and TrueGuide Synthetic sgRNA.
Once DNA fragments coding for CRISPR-Cas9 system components are cloned, the vector can be propagated within E. coli cells to generate sufficient quantities of your expression plasmid. We offer a variety of competent E. coli cells, selection of which depends upon the transformation method, and throughput of your experiment.
There are different methods to verify that your plasmid construct is correct (i.e., PCR, restriction digestion, or sequencing). The choice depends on whether you are interested in determining if the plasmid contains your DNA insert, is the insert in the right orientation, or does the insert have the correct sequence. We provide molecular biology tools to implement any analysis method you may choose.
Products for plasmid transformation and subsequent screening of clones | |
|---|---|
| For efficient transformation of constructed CRISPR plasmids you need competent E. coli cells: | |
.jpg) |
|
.jpg) | Isolate plasmid DNA in sufficient quantities and maximal purity for subsequent delivery to eukaryotic cells. Generate high yields of endotoxin-free plasmid (additional plasmid DNA isolation formats and sizes available). |
| Before proceeding into following steps you may wish to verify that your plasmid constructs are correct. Choose between different analysis methods: | |
.jpg) |
|
Accessory products
The CRISPR-Cas9 system simplifies genome editing and has great promise in broad applications such as stem cell engineering, gene therapy, tissue and animal disease models, and engineering disease-resistant transgenic plants.
Transfection is the process by which CRISPR-Cas9 DNA, mRNA, or protein systems are introduced into eukaryotic cells. Construct delivery techniques vary widely and include lipid nanoparticle–mediated transfection, viral delivery, and physical methods such as electroporation.
Products used for CRISPR construct delivery into eukaryotic cells | |
|---|---|
 |
|
Cell Culture Tools and Essentials:
- Sera—For your specific cell culture needs: from basic research to specialty assays
- Cell Culture Plastics—For optimum cell growth and consistency in cell culture
- Cell Culture Essentials—Find the tools and resources you need for successful cell culture
Whichever genome editing strategy you use, careful monitoring of the process will help you generate robust and reliable results. Start with accurate cell counts and viability determinations, then screen and validate the genotype of your cells. The genotyping technique of the mutant sequence depends on the type of mutation introduced through the CRISPR-generated edit. Here are the most common techniques:
- PCR amplification and gel electrophoresis for detection of larger Indels
- Mismatch-cleavage assay for Indel detection (T7 Endonuclease I cleavage assay)
- PCR amplification and restriction digestion for HDR analysis
- PCR amplification and cloning followed by Sanger sequencing
- PCR amplification and NGS
Products used to detect CRISPR-mediated genome modifications | |
|---|---|
.jpg) | Kits and reagents for creating clones carrying DNA fragments with sequences from genomic targets for subsequent Sanger sequencing |
.jpg) | Kit for rapid and quantitative measurements of CRISPR-Cas9 cleavage efficiency at your gene of interest |
 | Kits For isolating plasmid DNA and rapid, real-time DNA analysis using gel electrophoresis: |
| For accurate amplification of the target genomic regions, from crude or PCR-amplified samples:
|
.jpg) | Combine the reliability and performance you’ve come to expect from Applied Biosystems thermal cyclers with the flexible configuration and control features that fit how you work today |
| In as little as 1.5 hours, generate NGS libraries with the best coverage of all genomic regions to detect desired CRISPR-mediated events and search for potential off-target effects. |
.jpg)
CRISPR is routinely used for knockout, knock-in, or modulation of gene expression, and the effects can be measured using cell analysis techniques. Real-time PCR allows monitoring changes in the expression at gene level, for example when non-sense mediated decay decreases transcript levels, while western blotting is used to view changes to protein expression in a cell population; flow cytometry provides the throughput for multiparameter analysis on vast numbers of individual cells. Imaging allows for direct analysis of changes in protein expression, compartmentalization, and cell morphology, while high-content analysis (HCA) provides automation for the imaging process with quantitative rigor.
Products for further CRISPR analysis and edited cell collection | |
|---|---|
.jpg) | For the expression analysis by real time PCR isolate transcript RNA from the edited cells and transcribe them into cDNA: |
.jpg) | For the differential gene expression detection, you may perform whole transcriptome sequencing. We recommend preparing cDNA libraries that enables strand-specific RNA sequencing on Illumina next-generation sequencing (NGS) systems |
.jpg) |
|
The CRISPR-Cas9 system is a two-component system, consisting of the target-specific CRISPR gRNA and Cas9 nuclease. For genome editing to be successful, both the Cas9 and gRNA need to be expressed together in the target cells. We offer the necessary tools for the design and engineering of target-specific gRNA and Cas9 nuclease expression plasmids enabling you to pursue different experimental strategies.
Products for CRISPR-Cas9 design
For designing your vector you may choose between two separate plasmids for Cas9 and gRNA, respectively, or a single plasmid for both genes:
- Plasmid for Cas nuclease expression obtained from open-access resources (e.g., Addgene, the nonprofit global plasmid repository)
- gRNA cloning vector obtained from open-access resources (e.g., Addgene)
- GeneArt CRISPR Nuclease vector for cloning Cas9 and gRNA together
All-in-one plasmid system
Transfect, enrich, screen, and publish by using our GeneArt CRISPR Nuclease Vector Kit. The CRISPR Nuclease system offers a ready-to-use, all-in-one expression vector system with a Cas9 nuclease expression cassette and a guide RNA cloning cassette for fast cloning of a target-specific crRNA. This system allows you to edit and engineer the genomic locus of your choice in a sequence-specific manner from a single plasmid. After relevant targets have been identified with GeneArt CRISPRs, the biologically-relevant mutations can be validated with GeneArt TALs to reduce potential off-targeting.
GeneArt CRISPR Nuclease Vector Kits
GeneArt CRISPR Nuclease Vector Kits are reporter vector systems for expression of the functional components needed for CRISPR-Cas genome editing. The kits make it easy to express noncoding guide RNA (including crRNA and tracrRNA), using a plasmid vector that also expresses Cas9 endonuclease.
The GeneArt CRISPR Nuclease Vector with OFP (orange fluorescent protein) for flow cytometry-based sorting of crRNA-expressing cell populations, whereas GeneArt CRISPR Nuclease Vector with CD4 enables bead-based enrichment of crRNA-expressing cells.
GeneArt CRISPR Nuclease Vectors. (A) GeneArt CRISPR Nuclease: OFP Reporter Plasmid map and features of GeneArt CRISPR Nuclease: OFP Reporter. The vector is supplied linearized between nucleotides 6,732 and 6,752, with 5 bp 5´ overhangs on each strand as indicated. (B) GeneArt CRISPR Nuclease: CD4 Enrichment Plasmid map and features of GeneArt CRISPR Nuclease: CD4 Enrichment. The vector is supplied linearized between nucleotides 7,336 and 7,355, with 5 bp 5´ overhangs on each strand as indicated. 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.
| For constructing your own plasmid for gRNA expression you may need: | |
|---|---|
| |
| For easy cloning of novel CRISPR nuclease sequences or subcloning work you may need: | |
|
|
| Custom CRISPR for every gene; we'll design your target and clone it for you. Ready-to-transfect. | Contact us |
Related products
Invitrogen TrueTag Donor DNA Kits can help you obtain up to 100% knocked-in cells using our predesigned and validated donor DNA templates.
Looking for our premium genome editing solutions?
Learn more about our first-class genome engineering products, including TrueCut Cas9 v2 and TrueGuide Synthetic sgRNA.
Once DNA fragments coding for CRISPR-Cas9 system components are cloned, the vector can be propagated within E. coli cells to generate sufficient quantities of your expression plasmid. We offer a variety of competent E. coli cells, selection of which depends upon the transformation method, and throughput of your experiment.
There are different methods to verify that your plasmid construct is correct (i.e., PCR, restriction digestion, or sequencing). The choice depends on whether you are interested in determining if the plasmid contains your DNA insert, is the insert in the right orientation, or does the insert have the correct sequence. We provide molecular biology tools to implement any analysis method you may choose.
Products for plasmid transformation and subsequent screening of clones | |
|---|---|
| For efficient transformation of constructed CRISPR plasmids you need competent E. coli cells: | |
|
|
| Isolate plasmid DNA in sufficient quantities and maximal purity for subsequent delivery to eukaryotic cells. Generate high yields of endotoxin-free plasmid (additional plasmid DNA isolation formats and sizes available). |
| Before proceeding into following steps you may wish to verify that your plasmid constructs are correct. Choose between different analysis methods: | |
|
|
Accessory products
The CRISPR-Cas9 system simplifies genome editing and has great promise in broad applications such as stem cell engineering, gene therapy, tissue and animal disease models, and engineering disease-resistant transgenic plants.
Transfection is the process by which CRISPR-Cas9 DNA, mRNA, or protein systems are introduced into eukaryotic cells. Construct delivery techniques vary widely and include lipid nanoparticle–mediated transfection, viral delivery, and physical methods such as electroporation.
Products used for CRISPR construct delivery into eukaryotic cells | |
|---|---|
|
|
Cell Culture Tools and Essentials:
- Sera—For your specific cell culture needs: from basic research to specialty assays
- Cell Culture Plastics—For optimum cell growth and consistency in cell culture
- Cell Culture Essentials—Find the tools and resources you need for successful cell culture
Whichever genome editing strategy you use, careful monitoring of the process will help you generate robust and reliable results. Start with accurate cell counts and viability determinations, then screen and validate the genotype of your cells. The genotyping technique of the mutant sequence depends on the type of mutation introduced through the CRISPR-generated edit. Here are the most common techniques:
- PCR amplification and gel electrophoresis for detection of larger Indels
- Mismatch-cleavage assay for Indel detection (T7 Endonuclease I cleavage assay)
- PCR amplification and restriction digestion for HDR analysis
- PCR amplification and cloning followed by Sanger sequencing
- PCR amplification and NGS
Products used to detect CRISPR-mediated genome modifications | |
|---|---|
| Kits and reagents for creating clones carrying DNA fragments with sequences from genomic targets for subsequent Sanger sequencing |
| Kit for rapid and quantitative measurements of CRISPR-Cas9 cleavage efficiency at your gene of interest |
| Kits For isolating plasmid DNA and rapid, real-time DNA analysis using gel electrophoresis: |
| For accurate amplification of the target genomic regions, from crude or PCR-amplified samples:
|
| Combine the reliability and performance you’ve come to expect from Applied Biosystems thermal cyclers with the flexible configuration and control features that fit how you work today |
| In as little as 1.5 hours, generate NGS libraries with the best coverage of all genomic regions to detect desired CRISPR-mediated events and search for potential off-target effects. |
CRISPR is routinely used for knockout, knock-in, or modulation of gene expression, and the effects can be measured using cell analysis techniques. Real-time PCR allows monitoring changes in the expression at gene level, for example when non-sense mediated decay decreases transcript levels, while western blotting is used to view changes to protein expression in a cell population; flow cytometry provides the throughput for multiparameter analysis on vast numbers of individual cells. Imaging allows for direct analysis of changes in protein expression, compartmentalization, and cell morphology, while high-content analysis (HCA) provides automation for the imaging process with quantitative rigor.
Products for further CRISPR analysis and edited cell collection | |
|---|---|
| For the expression analysis by real time PCR isolate transcript RNA from the edited cells and transcribe them into cDNA: |
| For the differential gene expression detection, you may perform whole transcriptome sequencing. We recommend preparing cDNA libraries that enables strand-specific RNA sequencing on Illumina next-generation sequencing (NGS) systems |
|
|
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Resources
Learning Center
Access genome editing application resources for more success as you plan and execute your experiments.
Simplified CRISPR-Cas9 Protocols
Getting started with CRISPR-Cas9? Need proven genome editing protocols? Check out our collection of step-by-step and cell-line specific recommendations.
FAQs
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CRISPR Genome Editing Brochure
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