CRISPR Products & Services
CRISPR-Cas9 is a tool used for genome editing that has been adapted from the bacteria’s innate immune response. Bacteria capture and store DNA fragments from invading viruses or DNA within a region of their genome termed the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) array. This region is continually transcribed, producing short CRISPR RNA (crRNA) guide sequences that, when they detect an invading virus or DNA whose sequence is complementary to it, bind and recruit the Cas9 (CRISPR-associated protein 9) nuclease, which then specifically cleaves the invading DNA. This results in degradation of the invading DNA, thereby protecting the cell.
This CRISPR-Cas9 system has also been modified for use in mammalian cells. By expressing the Cas9 nuclease in mammalian cells and by introducing a single guide RNA sequence (sgRNA) specific for our gene of interest, we can introduce double-stranded breaks into the cell’s genomic DNA to either knock-out or modify a gene of interest.
CRISPR Knockout
Cas9 (Streptococcus pyogenes CRISPR associated protein 9) is an endonuclease enzyme that, when recruited to a specific DNA sequence by the sgRNA (single guide RNA), introduces a double stranded break into the DNA. This double stranded break is most commonly repaired in mammalian cells through Non-Homologous End Joining (NHEJ).
NHEJ often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation or knock-out of the targeted gene. To knock-out your gene of interest, BPS Biosciences offers the following solutions:
Integrating sgRNA & Cas9 Knockout Lentivirus
Our Cas9 Lentiviruses are replication incompetent, HIV-based VSV-G pseudo-typed lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. These particles contain a CRISPR/Cas9 gene driven by an EF1a promoter, along with 4 validated sgRNA (single guide RNA) targeting your gene of interest, driven by a U6 promoter.
Our Lentiviruses arrive ready-to-transduce – no other components are required - and our integrating sgRNA & Cas9 Lentiviruses are the surest way to generate a knock-out cell line or cell pool quickly; knock-out efficiencies can be as high as 90%, depending on the gene and cell type.
Non-integrating sgRNA & Cas9 Knockout Lentivirus
Unlike our Integrating sgRNA & Cas9 Lentiviruses, our Non-integrating Lentiviruses are made with a mutated Integrase, resulting in only transient expression of the sgRNA and Cas9 in your target cell. Although this may result in lower knock-out efficacies (depending on your gene and cell type) compared to our Integrating lentiviruses, the Non-integrating Lentiviruses eliminate the risk of the lentivirus randomly integrating into a physiologically-relevant gene and minimizes any potential off-targeting that may occur if Cas9 is stably expressed. All our Lentiviruses are validated and arrive ready-to-transduce.
These cell pools already constitutively express Cas9. To knock-out your gene of interest, you only need to deliver your gene-specific sgRNA via electroporation or lentiviral transduction. Affordably priced, these can be a cost-effective platform for setting up your own knock-out experiments or screens.
Cas9 Protein
Our Cas9 protein can be utilized in in vitro cleavage assays to compare and identify sgRNAs with the highest cutting efficiencies.
With BPS Bioscience’s custom cell line development services, our team of highly experienced scientists can generate custom knock-out cell lines in more than 70 different cell types using CRISPR-Cas9 licensed technology, targeting whichever gene(s) of interest you’re interested in. The development process is comprised of separate milestones where data is provided after each milestone completion. Each project is customized for the desired deliverables through working directly with you. Contact us to learn more about what we can offer.
CRISPR Knock-In
When Cas9 generates a double stranded break in the cell’s genomic DNA, the mammalian cell repairs that break through either Non-Homologous End Joining (NHEJ) or Homology Directed Repair (HDR). When the cell undergoes HDR, we can provide an alternative template, either a ssDNA oligo or a plasmid donor, that contains whichever genetic mutations (mutations or tags) we want to introduce flanked by homology arms, and the cell will use that as a template to copy and incorporate the changes into the genomic DNA.
With BPS Bioscience’s custom cell line development services, our team of highly experienced scientists can generate custom knock-in cell lines in more than 70 different cell types using CRISPR-Cas9 licensed technology, targeting whichever gene(s) of interest you’re interested in. The development process is comprised of separate milestones where data is provided after each milestone completion. Each project is customized for the desired deliverables through working directly with you. Contact us to learn more about what we can offer.
CRISPR Activation
The CRISPR Synergistic Activation Mediator (SAM) system is used to induce transcriptional activation and expression of any gene of interest. The SAM system consists of 3 components: (1) a mutated dCas9 (Streptococcus pyogenes CRISPR associated protein 9), lacking any endonuclease activity, fused to VP64, a transcriptional activator; (2) P65 (Transcription Factor p65, or Nuclear Factor NF-κ-B p65) and HSF1 (Heat Shock Factor 1) fused with an MS2 tag; and (3) the MS2-tagged sgRNA targeting the promoter region of the gene of interest.
Once the sgRNA-MS2 targets the promoter region of the gene of interest, dCas9-VP64 and MS2-P65-HSF1 are recruited to the site in the genomic DNA and begin transcription, inducing expression of the desired gene. Induction can be more than a hundred-fold, depending on the gene.
To utilize the SAM system to induce expression of your gene of interest, BPS Biosciences offers the following solutions:
CRISPR Activation (CRISPRa) Cell Lines
These cell lines stably express all the required components for activating your gene of interest except for the sgRNA-MS2. These cells express a mutated dCas9 (lacking any endonuclease activity) fused to VP64, a transcriptional activator, and P65 (Transcription Factor p65, or Nuclear Factor NF-κ-B p65) and HSF1 (Heat Shock Factor 1) fused with an MS2 tag.
When these cells are transduced with an MS2-tagged sgRNA targeting the promoter region of your gene of interest, the dCas9-VP64 and MS2-P65-HSF1 are recruited to the site in the genomic DNA and begin transcription, inducing expression of your desired gene. The gene-specific sgRNA-MS2 can be delivered via lentiviral transduction or electroporation. These cell lines can be a convenient and cost-effective platform for setting up your own activating experiments or screens without needing to first select for single cell clones expressing dCas9-VP64 and MS2-P65-HSF1, saving valuable time.
Integrating dCas9-VP64 and MS2-P65-HSF1 Lentiviruses
Our Cas9 Lentiviruses are replication incompetent, HIV-based VSV-G pseudo-typed lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. These particles contain the dCas9-VP64 (with Blasticidin resistance) and MS2-P65-HSF1 (with Hygromycin resistance) genes and can be used, in conjunction with the sgRNA-MS2 Activating Lentivirus, to generate your own CRISPR Activation (CRISPRa) cell lines.
Integrating sgRNA-MS2 Activating Lentivirus
These Lentivirus particles contain 4 validated sgRNA (single guide RNA) targeting the promoter region of your gene of interest, fused to MS2 and driven by a U6 promoter. These Lentiviruses arrive ready-to-transduce into your dCas9-VP64 and P65-HSF1-MS2 expressing cell lines to stably activate expression of your gene of interest.
While the Integrating sgRNA-MS2 Lentiviruses can be used to generate cell pools or lines stably inducing expression of your gene of interest, the sgRNA-MS2 plasmids can be transfected or electroporated into your cells for either transient or stable (following drug selection) activation. This also provides you with a virus-free option, depending on your laboratory environment or preferences.
Custom CRISPR Activation (CRISPRa) Services
With BPS Bioscience’s custom cell line development services, our team of highly experienced scientists can generate custom CRISPR activation cell lines in more than 70 different cell types using CRISPR-Cas9 licensed technology, targeting whichever gene(s) of interest you’re interested in. The development process is comprised of separate milestones where data is provided after each milestone completion. Each project is customized for the desired deliverables through working directly with you. Contact us to learn more about what we can offer.
CRISPR Screens
CRISPR Kinase Knockout Library, Array or Pool (Coming soon!)
BPS Bioscience is building a CRISPR Knockout library targeting human kinases, available in both array and pooled formats. Our Integrating Lentiviruses are shipped ready-to-transduce into almost all types of mammalian cells, including primary and non-dividing cells. Each format includes 4 sgRNA per gene, in addition to the proper controls. Our array format comes ready-to-use, delivered as one gene-per-well, without the need for any high-throughput screening platforms or bioinformatic analysis. Our pooled format is delivered at high titer, and provides a cheaper alternative, depending on your screening capabilities.
With BPS Bioscience’s custom cell line development services, our team of highly experienced scientists can generate CRISPR libraries using CRISPR-Cas9 licensed technology, targeting whichever gene(s) of interest you’re interested in. The development process is comprised of separate milestones where data is provided after each milestone completion. Each project is customized for the desired deliverables through working directly with you. Contact us to learn more about what we can offer.
