CRISPR Genome Editing Solutions

VectorBuilder offers a variety of CRISPR products and services for in vitro and in vivo genome editing experiments at unbeatable prices and with rapid turnaround. Our CRISPR offerings range from off-the-shelf reagents that are ready for transfection or transduction to custom-made CRISPR vectors, available in multiple delivery formats (i.e. plasmid, virus, RNA). We can package all major viral types (i.e. lentivirus, AAV, adenovirus) to deliver your CRISPR components efficiently into difficult-to-transfect cells. Additionally, we are specialized in building high-quality CRISPR libraries for knockout, gene activation, gene inhibition and other CRISPR screens. Our uniquely designed and well validated whole genome dual-gRNA knockout libraries are powerful tools for gene functional screens in human and mouse.

Additionally, our online vector design platform features a free and user-friendly CRISPR design tool that allows you to design CRISPR vectors with high targeting efficiency. VectorBuilder has created whole-genome gRNA databases that contain optimized gRNA sequences for common species. When you design CRISPR vectors on VectorBuilder, you will have the option to search for your target gene in our database. Upon entering your target gene, you will see detailed information on all the available guide RNA designs in our database corresponding to your gene. This feature enables you to select gRNAs with high specificity and minimal off-target effects, thereby providing you with advantages offered by popular gRNA design tools or software.

With our extensive experience in molecular cloning, gene delivery and gene targeting, VectorBuilder provides one-stop solutions for all your CRISPR gene editing needs.

Click the links below to view the detailed description of our CRISPR offerings:
Highlights of VectorBuilder's CRISPR products & services
  • Highly intuitive online vector design platform with whole-genome gRNA database implemented for easy and quick CRISPR design
  • Rich collection of vector backbones (i.e. regular plasmid, lentivirus, AAV, adenovirus, piggyBac) and vector components (i.e. promoters, Cas9 variants, fluorescent and drug-selection markers)
  • CRISPR components available in a variety of delivery formats (i.e. CRISPR/Cas9 plasmid, CRISPR/Cas9 virus, Cas9 mRNA + gRNA, Cas9-gRNA RNP complex)
  • Versatile CRISPR library construction options
  • Robust quality, fast turnaround and competitive pricing
  • Powerful technical support for experimental design, data analysis and troubleshooting
CRISPR-mediated genome editing

The conventional use of CRISPR system contains two components: Cas9 protein and guide RNA (gRNA). The most commonly used Cas9 is engineered from Streptococcus pyogenes (a.k.a. SpCas9 or hCas9). It is an RNA-guided DNA nuclease which can generate double-stranded breaks (DSBs) at target sites (Figure 1). Another widely used Cas9 variant, Cas9 “nickase” (e.g. Cas9(D10A)), generates single-stranded cuts in DNA. If Cas9 nickase is used in conjunction with two gRNAs targeting the two opposite strands flanking a single target region, the nickase will generate two single-stranded cuts, one on each strand, resulting in DSBs at the target region. Once DSBs are generated by the CRISPR system, cells activate the nonhomologous end-joining (NHEJ) pathway to repair the DNA breaks, which usually results in small random deletions, or more rarely insertions and base substitutions. When these mutations disrupt a protein-coding region (e.g. a deletion causing a frameshift), they may lead to functional gene knockout. Alternatively, and less efficiently, DSBs can be repaired via the homology-directed repair (HDR) pathway, when an exogenous donor DNA template is co-introduced with the CRISPR components. This can result in replacement of the target genomic DNA sequence with donor sequence, generating precise sequence changes such as point mutations or knockin of the donor DNA sequence at the target site. The donor DNA template can be a single-stranded oligo nucleotide (ssODN) or a dsDNA fragment (usually linearized plasmid DNA). ssODNs are suitable for introducing point mutations or small tag insertions, while dsDNA fragments are widely used to introduce large fragment knockins.

Figure 1. Mechanism of CRISPR-induced DNA repair.

CRISPR delivery approaches

CRISPR components can be introduced into mammalian cells via different approaches (Figure 2), including:

  • gRNA and Cas9 plasmid
  • gRNA and Cas9 virus (i.e. lentivirus, AAV, adenovirus, etc.)
  • Mixture of gRNA and Cas9 mRNA
  • Preformed ribonucleoprotein (RNP) between gRNA and Cas9 protein
Figure 2. Four methods for delivering CRISPR components into cells.

The table below lists key advantages and disadvantages for each delivery approach and can be used as a reference to help you decide the most suitable approach for your experiments:

Delivery Approach Advantages Disadvantages
gRNA and Cas9 plasmid
  • Delivered by chemical transfection or electroporation which is simple and low-cost.
  • Able to achieve high-level expression of Cas9 protein and gRNA for a few days.
  • Cas9 and gRNA can be delivered in all-in-one plasmid, therefore circumventing the need for transfecting multiple components.
  • Plasmids as reagents can be generated and regenerated very cheaply and in very large quantities, making them suitable for CRISPR experiments that require a lot of reagents.
  • Efficiency of plasmid transfection varies widely between cell types.
  • Dependent on cell-type specific promoter activity.
  • Largely limited to in vitro applications.
  • Risk of random integration of plasmid DNA into the host genome.
  • Risk of off-target effects due to high-levels of Cas9 and gRNA expression.
gRNA and Cas9 virus
  • Suitable for genome editing applications in difficult-to-transfect cells.  
  • Cas9 and gRNA can be delivered in a single all-in-one viral vector, therefore circumventing the need for transduction of multiple components.
  • Can achieve prolonged or stable expression of Cas9 protein and gRNA.
  • Requires virus packaging which is technically challenging and time-consuming, unless outsourced to VectorBuilder.
  • Dependent on cell-type specific promoter activity.
  • Elevated risk of insertional mutagenesis.
  • Risk of off-target effects due to prolonged Cas9 expression.
Mixture of gRNA and Cas9 mRNA 
  • Delivered by chemical transfection or electroporation which is simple.
  • Rapid genome editing as no transcription is needed for Cas9 and gRNA expression.
  • Independent of cell-type specific promoter activity.
  • No risk of random insertion into host genome.
  • Editing activity occurs only transiently, after which it drops off quickly as the transcripts are degraded inside the host cells.  
Preformed gRNA-Cas9 RNP complex
  • Can be delivered by electroporation, making it suitable for difficult-to-transfect cells.
  • Rapid editing as neither transcription nor translation is needed for Cas9 and gRNA expression.
  • Independent of cell-type specific promoter activity.
  • No risk of random insertion into host genome.
  • Electroporation-based RNP delivery requires expensive hardware and consumables.
  • Editing activity occurs as a rapid pulse, after which it drops off quickly as gRNA-Cas9 RNP is degraded inside the host cells.
  • Limited by the availability of purified Cas9 protein.
Our CRISPR offerings
Custom CRISPR vectors

CRISPR-mediated genome editing requires target cells to co-express Cas9 and target site-specific gRNA at the same time. Using our highly intuitive online vector design platform, you can choose from over 30 vector backbones (non-viral, viral or transposon) and unlimited combinations of vector components (promoters, Cas9 variants, fluorescent and drug-selection markers) to express Cas9 and your gRNA(s) in different manners. Our gRNA database allows you to easily pick appropriate guide sequence for your target genes without looking for and analyzing the target sites on your own. We offer all-in-one as well as dual vector systems for Cas9 and gRNA expression in regular plasmid, lentivirus, AAV, adenovirus and PiggyBac backbones.

Our comprehensive collection of CRISPR vectors also includes gene targeting donor vectors serving as DNA templates to guide precise sequence changes such as point mutations and large fragment knockins at target sites through HDR. Additionally, we offer CRISPRa and CRISPRi vectors for achieving transcriptional activation or inhibition of target genes, respectively, within their endogenous loci. We can design and construct vectors for dCas9-SAM-based or dCas9-SunTag-based CRISPR activation and dCas9-KRAB or dCas9-KRAB-MeCP2 based CRISPR inhibition applications. 

Plasmid DNA preparation, RNA preparation and virus packaging can be purchased as downstream services when adding CRISPR vectors into your shopping cart.

Our collection of CRISPR vectors include:

Note: AAV has a cargo capacity of 4.7 kb. The commonly used SpCas9 derived from Streptococcus pyogenes is 4.2 kb, which can barely fit into AAVs when combined with other components such as a promoter, the polyA signal sequence, and the gRNA expression cassette. Our standard AAV CRISPR system therefore utilizes the shorter SaCas9 (3.2 kb) derived from Staphylococcus aureus. Please note that the PAM sequence recognized by SaCas9 is NNGRR or NNGRRT (preferred), whereas the PAM for SpCas9 is NGG. We can also construct AAV SpCas9 vectors upon request.

Premade CRISPR vectors

In addition to custom CRISPR/Cas9 vectors, VectorBuilder also offers a panel of premade Cas9 vectors, control gRNA vectors and helper vectors for CRISPRa and CRISPRi. Our off-the-shelf vectors are available for immediate shipment as E. coli stocks. Plasmid DNA preparation, RNA preparation and virus packaging can be purchased as downstream services upon adding premade vectors into your shopping cart.

Our collection of premade CRISPR vectors include:

Vector System Vector Name Vector ID Price (USD)
hCas9 expression vector (regular plasmid) pRP[Exp]-
mCherry/Hygro-CBh>hCas9
VB170315-1076pcv $159
hCas9 expression vector (lentivirus) pLV[Exp]-
CBh>hCas9:T2A:Hygro
VB160923-1033trt $99
SaCas9 expression vector (AAV) pAAV[Exp]-
CMV>SaCas9
VB180125-1089ffa $159
hCas9 expression vector (adenovirus) pAV[Exp]-CBh>hCas9 VB180820-1032mbq $159
hCas9 expression vector (piggyBac) pPB[Exp]-mCherry/Hygro-CBh>hCas9 VB181219-1015jaf $159
Scramble gRNA control vector (regular plasmid) pRP[gRNA]-EGFP/Puro-U6>Scramble_gRNA VB171211-1212bte $99
Scramble gRNA control vector (lentivirus) pLV[gRNA]-EGFP/Puro-U6>Scramble_gRNA VB160602-1127urj $99
Scramble SagRNA control vector (AAV) pAAV[gRNA]-EGFP/Puro-U6>Scramble_SagRNA1 VB181225-1325hfs $99
Scramble gRNA control vector (adenovirus) pAV[gRNA]-EGFP-U6>Scramble_gRNA1 VB180416-1105sbw $239
Scramble gRNA control vector (piggyBac) pPB[gRNA]-EGFP/Puro-U6>Scramble_gRNA1 VB181225-1329xyy $99
hCas9 and scramble gRNA coexpression vector (regular plasmid) pRP[CRISPR]-EGFP/Puro-hCas9-U6>Scramble_gRNA1 VB180725-1096pqd $99
hCas9 and scramble gRNA coexpression vector (lentivirus) pLV[CRISPR]-hCas9/Puro-U6>Scramble_gRNA1 VB180522-1197bhe $99
SaCas9 and scramble SagRNA coexpression vector (AAV) pAAV[CRISPR]-SaCas9-U6>Scramble_SagRNA VB170425-1003ytn $95
hCas9 and scramble gRNA coexpression vector (adenovirus) pAV[CRISPR]-hCas9/EGFP-U6>Scramble_gRNA1 VB181225-1306tra $239
dCas9-SAM activator MS2/P65/HSF1 expression vector (lentivirus) pLV[Exp]-EF1A>MS2/P65/HSF1/Hygro VB170424-1011udy $159
dCas9-SAM activator dCas9/VP64 expression vector (lentivirus) pLV[Exp]-EF1A>dCas9/VP64/Bsd VB170424-1009jup $159
dCas9-SAM scramble msgRNA control vector (lentivirus) pLV[msgRNA]-EGFP/Puro-U6>Scramble_gRNA VB180124-1069tzr $99
dCas9-KRAB-MeCP2 expression vector (lentivirus) pLV[Exp]-CBh>dCas9/
KRAB/MeCP2:T2A:Hygro
VB190101-1021vwr $159
CRISPR virus

Lentivirus, AAV and adenovirus are widely used to deliver CRISPR components into mammalian cells. VectorBuilder offers premium-quality virus packaging services for lentivirus, AAV and adenovirus for achieving highly efficient CRISPR targeting in difficult-to-transfect cells. Our proprietary technologies and reagents have greatly improved virus packaging protocols in terms of titer, purity, viability and consistency. Our packaging protocols are also optimized for the viral vector systems used in our vector construction services. As a result, we have a growing base of highly satisfied customers who come back to us again and again for their cloning and virus packaging needs.

Price, turnaround and scales of our lentivirus packaging services:

Scale Application Titer & Volume Price (USD) Turnaround
Pilot Cell culture >108 TU/ml, 10x25 ul, HBSS buffer $399 8-16 days
Medium Cell culture >108 TU/ml, 10x100 ul, HBSS buffer $599 8-16 days
Large Cell culture >109 TU/ml, 10x100 ul, HBSS buffer $999 8-16 days
Ultra-purified medium Cell culture and in vivo >109 TU/ml, 10x50 ul, HBSS buffer $1199 8-16 days
Ultra-purified large Cell culture and in vivo >109 TU/ml, 10x100 ul, HBSS buffer $1499 8-16 days

Price, turnaround and scales of our AAV packaging services:

Scale Application Titer & Volume Price (USD) Turnaround
Pilot Cell culture >2x1011 GC/ml, 10x25 ul, PBS buffer $399 10-20 days
Medium Cell culture >2x1011 GC/ml, 10x100 ul, PBS buffer $549 10-20 days
Large Cell culture >2x1012 GC/ml, 10x100 ul, PBS buffer $999 10-20 days
Ultra-purified pilot Cell culture and in vivo >1013 GC/ml, 4x25 ul, PBS buffer $1149 20-30 days
Ultra-purified medium Cell culture and in vivo >1013 GC/ml, 10x50 ul, PBS buffer $1649 20-30 days
Ultra-purified large Cell culture and in vivo >1013 GC/ml, 10x100 ul, PBS buffer $2549 20-30 days

Price, turnaround and scales of our adenovirus packaging services:

Scale Application Titer & Volume Price (USD) Turnaround
Pilot Cell culture >1010 PFU/ml, 10x25 ul, HBSS buffer $399 20-32 days
Medium Cell culture >1010 PFU/ml, 10x100 ul, HBSS buffer $599 20-32 days
Large Cell culture >1011 PFU/ml, 10x100 ul, HBSS buffer $999 20-32 days
Ultra-purified large Cell culture and in vivo >1012 VP/ml, 10x100 ul, GTS buffer $1499 22-36 days

Note:

a. Estimated turnaround is the time from production initiation to completion. It does not include waiting time for any customer-supplied materials (e.g. template DNA), QC of such materials, and transit time for shipping final deliverables to the customer.

b. Our titer guarantee only applies to vectors for which the region being packaged into virus does not exceed the cargo capacity of the virus. For lentivirus, it is 9.2 kb (from Δ5’ LTR to ΔU3/3’ LTR). For AAV, it is 4.7 kb (from 5' ITR to 3' ITR). For adenovirus, it is 38.7 kb (from 5' ITR to 3' ITR).

c. We are not able to guarantee titer for the following vectors:

  • vectors containing sequences that could adversely affect the packaging process such as toxic genes (e.g. proapoptotic genes), genes that compromise the integrity of packaging cells or virus (e.g. membrane proteins that cause cell aggregation), and sequences prone to rearrangements or secondary structures (e.g. repetitive or highly GC-rich sequences);
  • customer-supplied vectors as we have no control over their quality.

Click here to view detailed information on our virus packaging services

RNA preparation for Cas9 mRNA and gRNA

VectorBuilder can provide transfection-ready and microinjection-ready Cas9 mRNA and gRNA specifically designed against user-selected target sites for easy RNA-based delivery of CRISPR components into mammalian cells. Cas9 mRNA is available for both wildtype hCas9 nuclease and Cas9 nickase (Cas9(D10A)). We follow the algorithm developed in Feng Zhang’s lab to calculate specificity score and apply a set of empirical rules to design the optimal gRNA for targeting user-selected genes/sites.

Price and turnaround of our CRISPR RNA products:

Reagent Concentration & Volume Price (USD) Turnaround
hCas9 mRNA >500 ng/ul, 25 ul, nuclease-free water, sterile $349 3-5 days
Cas9(D10A) mRNA >500 ng/ul, 25 ul, nuclease-free water, sterile $349 3-5 days
Custom gRNA* >500 ng/ul, 25 ul, nuclease-free water, sterile $349 3-5 days

* Construction of gRNA in vitro transcription vector has an additional $99 charge and 7-14 days turnaround.

Cas9 protein

VectorBuilder offers purified wildtype Streptococcus pyogenes Cas9 protein (SpCas9) and the Cas9 nickase (Cas9(D10A)) for preparing preformed Cas9-gRNA RNP to deliver CRISPR components into mammalian cells. The RNP-mediated approach can be used to deliver your desired gRNA in conjunction with Cas9 protein. Additionally, preformed RNPs can also be delivered together with donor templates (ssODN or dsDNA) to induce HDR-mediated DNA repair at CRISPR target sites for precise genome editing.

Price and turnaround of our Cas9 protein products:

Reagent Concentration & Volume Price (USD) Turnaround
SpCas9 protein 100 ug $599 5-7 days
SpCas9(D10A) nickase protein 100 ug $599 5-7 days
Donor DNA for precise genome editing

VectorBuilder offers donor DNA templates in the form of ssODN or dsDNA from linearized plasmid for guiding HDR-based DNA repair to introduce precise DNA sequence changes at CRISPR cleavage sites. The introduced changes include point mutations and small or large fragment knockins. While ssODNs are utilized for inserting short DNA sequences (< 60 bp) such as small tags or restriction enzyme sites at target sites, dsDNA donors are utilized for targeted knockin of larger sequences (up to 4-5 kb) such as fluorescent tags and other reporters.

Price and turnaround of our donor DNA products:

Reagent Price (USD) Turnaround
ssODN (normally 120-200 bp) $349 2-3 weeks
Pooled CRISPR libraries

Pooled CRISPR libraries are powerful tools for performing large-scale functional screens of coding or noncoding regions involved in particular pathways, diseases, cell responses to drug treatment, developmental processes, gene regulation, etc. VectorBuilder specializes in the custom design and construction of a variety of pooled libraries for commonly used CRISPR knockout libraries for functional screens of coding genes, CRISPRa libraries for gain-of-function screens of coding genes or regulatory function screens of noncoding regions and CRISPRi libraries for loss-of-function screens of coding genes or for screening regulatory function of noncoding regions. Additionally, we can also build CRISPR barcode libraries for single cell-based screens and other pooled libraries utilizing CRISPR technologies.

We can deliver your library as an E. coli stock, plasmid DNA pool, or packaged virus, depending on your needs. Our custom libraries are fully validated by next-generation sequencing so that you know exactly what you get.

Click here for detailed information on our library construction services

In addition to custom pooled CRISPR libraries, we also offer premade dual-gRNA lentivirus libraries for whole-genome knockout screens in human and mouse. Dual-gRNA libraries are far more powerful than single-gRNA libraries for knockout screens because the introduction of large deletions by these libraries can have much higher efficiencies in generating loss-of-function mutations. The human and mouse dual-gRNA libraries are made in the form of ready-to-use high-titer pooled lentivirus targeting 20,048 human and 20,493 mouse genes, respectively. Wherever possible, each gene is targeted redundantly by 4-6 different gRNA pairs in separate vectors.

Highlights of our premade dual-gRNA CRISPR libraries:

  • Unique dual-gRNA lentiviral vector design
  • Whole-genome and high-coverage targeting
  • Validation of library quality by NGS
  • High uniformity
  • Available as ready-to-use high-titer lentivirus
  • Dual EGFP/Puro marker for efficient and versatile selection or tracking of positively transduced cells

Price and turnaround of our premade dual-gRNA libraries:

Product name No. of genes No. of gRNA pairs Scale* Catalog No. Price (USD)
Human Whole-Genome Dual-gRNA Lentivirus Library 20,048 91,926

Medium

(>1.0x108 TU/ml, 1 ml)

LVM(Lib190505-1046fgb) $7,999
$3,999

Plus

(>1.0x108 TU/ml, 5 ml)

LV5M(Lib190505-1046fgb) $16,999
$8,499
Mouse Whole-Genome Dual-gRNA Lentivirus Library 20,493 90,344

Medium

(>1.0x108 TU/ml, 1 ml)

LVM(Lib190505-1050kpm) $7,999
$3,999

Plus

(>1.0x108 TU/ml, 5 ml)

LV5M(Lib190505-1050kpm) $16,999
$8,499

Click here for detailed information on our premade dual-gRNA CRISPR libraries

CRISPR solutions

The versatility of the CRISPR system makes it suitable for a wide variety of genome editing applications in mammalian cells. Our technical team with extensive experience in molecular biology applications, can provide complete solutions for all your CRISPR genome editing projects from experimental design to generation of the ready-for-use reagents. We can help you with the common CRISPR applications listed below, and we can work with you on any novel CRISPR application. Send us a design request immediately to get a free service proposal!

  • Gene knockout through gene disruption
    • Random frameshift resulting from NHEJ DNA repair at single cut site
    • Exon deletion between two cut sites
  • Introduction of point mutation
  • Fragment knockin
    • Small tag insertion (< 60 bp)
    • Large fragment insertion (up to 4-5 kb)
  • CRISPR gene activation
  • CRISPR gene inhibition
  • CRISPR libraries (KO, CRISPRa, CRISPRi, barcoding, etc.)
  • Stable cell line generation
    • Cas9-expressing cell line
    • iPSC genome editing
    • Cancer cell genome editing

Click here for a detailed information on our stable cell line generation services

VectorBuilder's online “Learning Center” under "Resources" contains rich educational resources to facilitate you to successfully plan, execute and troubleshoot your CRISPR experiments.

Click here to read guides on CRISPR vector systems

Click here to read guides on CRISPR components

Click here to read CRISPR-related FAQs