Custom CRISPR Multiplex sgRNA Vectors

Interested in knocking out multiple genes or recruiting transcriptional regulators using a combination of sgRNAs? abm’s CRISPR multiplex gRNA system allows for optimal expression of multiple gRNAs from alternating U6 and H1 RNA pol III promoters on a single lentiviral vector. gRNAs may be designed for use with any Cas9 variant, which is expressed from a separate vector (click here to see abm’s full list of Cas9-expressing vectors and viruses). Just tell us what you want to accomplish, and we’ll do the sgRNA design, cloning, and lentiviral packaging for you.



Additional Resources:

Service Details

sgRNA Service (for spCas9)No. of sgRNAsQuantityCat. No.Price
Custom CRISPR Dual sgRNA Lentivector (for spCas9) 2 1.0 ug C420 $395.00
Custom CRISPR Triple sgRNA Lentivector (for spCas9) 3 1.0 ug C421 $595.00
Custom CRISPR Quad sgRNA Lentivector (for spCas9) 4 1.0 ug C422 $795.00
Custom CRISPR Multiplex sgRNA Lentivector (for spCas9) >4 1.0 ug C423 Inquire
sgRNA Service (for saCas9)No. of sgRNAsQuantityCAT. NO.PRICE
Custom CRISPR Dual sgRNA Lentivector (for saCas9) 2 1.0 ug C511 $395.00
Custom CRISPR Triple sgRNA Lentivector (for saCas9) 3 1.0 ug C512 $595.00
Custom CRISPR Quad sgRNA Lentivector (for saCas9) 4 1.0 ug C513 $795.00
Custom CRISPR Multiplex sgRNA Lentivector (for saCas9) >4 1.0 ug C514 Inquire
Packaging OptionsVolumeCAT. NO.PRICE
Custom Recombinant Lentivirus (106 IU/ml) 10 ml LV002 $650.00
High Titer Custom Recombinant Lentivirus (107 IU/ml) 1.0 ml LV002-a $850.00
High Titer Custom Recombinant Lentivirus (108 IU/ml) 550 ul LV002-b $1100.00
High Titer Custom Recombinant Lentivirus (109 IU/ml) 100 ul LV002-c $1350.00
High Titer Custom Recombinant Lentivirus (1010 IU/ml) 100 ul LV002-d $1850.00

Additional Info

Applications
  1. Multi-target genomic mutagenesis:
    • Target multiple sites at a single locus or target multiple different loci simultaneously
    • Convenient for use with Cas9 nickase to increase specificity and reduce off-target effects. Easily co-express multiple gRNAs simultaneously.
    • sgRNAs can be designed for use with any Cas9 (saCas9, spCas9, dCas9-SAM, dCas9-KRAB, etc.)
  2. Simultaneous activation, repression, knockout of multiple genomic targets:
    • Activation using dCas9-SAM
    • Repression using dCas9-KRAB
    • Knockout using spCas9 or saCas9

Workflow

* The vector for our CRISPR Multiplex sgRNA lentivectors is pLenti-Multi-gRNA-PGK-Neo.

FAQs

What promoter(s) do your sgRNA and All-in-One lentivectors have?
sgRNAs are expressed from the U6 promoter and All-in-One lentivectors and lentiviruses express Cas9 from the SFFV promoter.
What is the approximate time frame to generate CRISPR lentiviral constructs?
The typical lead time is around 1-3 weeks for CRISPR lentivectors and 4-6 weeks for lentiviruses. Please inquire for a more accurate lead time.
How should I store my lentivirus?
Aliquots should be made for the lentivirus and stored at -70 degrees Celsius.
If a high-titer custom lentivirus was ordered, do you do the lentiviral titration after a freeze-thaw or straight after production before the freeze-down step?
We do the titration after aliquoting and freeze-down. Thus, the titer should be accurate for the customer after they thaw the finished product for the first time.
Which virus has your lentivirus expression system been derived from? Is it HIV?
Our lentivirus expression system is derived from Human HIV-1 Virus. It employs third generation self-inactivating recombinant lentiviral vectors with enhanced biosafety features and minimal relation to wild-type Human HIV-1 Virus.
How do you verify the titer?
We use our LV900 series - qPCR Lentivirus Titration(Titer) Kit. This kit quantifies a proprietary region of the lentiviral 5’-LTR.
What is the packaging capacity for Lentivirus?
The maximum insert size is <9 kb between the 5'LTR to 3'LTR.

Citations

01 Kang, Y. J. et al. "Regulation of NKT cell-mediated immune responses to tumours and liver inflammation by mitochondrial PGAM5-Drp1 signalling." Nat. Commun. (2015) 6:8371 doi: 10.1038/ncomms9371
02 Jiang, G. et al. "Isorhapontigenin (ISO) inhibits invasive bladder cancer (BC) formation in vivo and human BC invasion in vitro by targeting STAT1/FOXO1 Axis." Cancer Prev Res. Published Online First April 14, 2016.doi: 10.1158/1940-6207.CAPR-15-0338
03 Okugawa, Y. et al. "Clinical significance of SNORA42 as an oncogene and a prognostic biomarker in colorectal cancer." Gut (2015)doi:10.1136/gutjnl-2015-309359