Cat.No. : CSC-RT2699
Host Cell: Jurkat Target Gene: CD47
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
| Cat. No. | CSC-RT2699 |
| Description | This cell is a stable cell line with a homozygous knockout of human CD47 using CRISPR/Cas9. |
| Target Gene | CD47 |
| Host Cell | Jurkat |
| Host Cell Species | Homo sapiens (Human) |
| Size Form | 1 vial (10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid nirtogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
CD47 is a ubiquitously expressed cell surface glycoprotein that has been implicated in immune evasion. It interacts with the inhibitory receptor signal regulatory protein α (SIRPα), which is primarily expressed on myeloid cells and normally functions to limit effector functions such as phagocytosis and immune cell homeostasis. CD47/SIRPα antagonists, often referred to as "macrophage checkpoint" inhibitors, are being developed as cancer interventions. SRF231 is an investigational fully human IgG4 anti-CD47 antibody currently being evaluated in clinical trials. The studies here demonstrate that SRF231 binds CD47 and disrupts the CD47/SIRPα interaction without causing hemagglutination or erythrophagocytosis. SRF231 exerts antitumor activity in vitro through phagocytosis and cell death in a manner that is dependent on activation of the Fc-gamma receptor (FcγR) CD32a. Through its Fc domain, SRF231 binds to macrophage-derived CD32a and plays a dual role, initiating FcγR-mediated phagocytosis of cancer cells and acting as a scaffold to drive CD47-mediated death signals into tumor cells.
In this study, SRF231 specifically bound to CD47, as shown by surface staining of WT Jurkat cells expressing CD47, but not to CD47 knockout Jurkat cells (Figure 1A). Multiple human tumor cell lines of the hematopoietic lineage were sensitive to SRF231-mediated phagocytosis by primary human macrophages (Figure 1B). In addition, a significant enhancement of phagocytosis of primary AML bone marrow cells was observed in the presence of SRF231 (Figure 1C). As expected, SRF231 synergized with anti-CD20 antibodies, resulting in enhanced antibody-dependent cellular phagocytosis (ADCP) of the malignant B-cell lymphoma cell lines Raji and SU-DHL-4 (Figure 1D).
Figure 1. SRF231 binds to human CD47 and induces phagocytosis of tumor cells in vitro. (Peluso M O, et al., 2020)
A: The knockout cell product is validated by PCR amplification and Sanger Sequencing to confirm the mutation at the genomic level. Please find the detailed mutation info in the datasheet.
A: Single clonal cell.
A: No. This knockout cell product is generated using the CRISPR/Cas9 system to induce small insertions or deletions (indels) resulting in frameshift mutations. Although these frameshift mutations typically disrupt the coding gene, there is a possibility that the non-functional transcript may still be transcribed. Consequently, this could potentially yield misleading results when analyzed by RT-qPCR.
A: The cell line should be stored in liquid nitrogen for long-term preservation.
A: For most cases, we often keep at least 2 clones with different frameshift mutations. Please feel free to contact us to check if there are additional available clones.
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I've been using the Human CD47 Knockout Cell Line-Jurkat for several months now. This cell line has significantly simplified my workflow and provided consistent, reproducible results.
The Human CD47 Knockout Cell Line has enabled me to conduct more precise functional assays, revealing novel insights into the role of CD47 in cell signaling pathways.
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