The CD22 Lentiviruses are replication incompetent, HIV based, VSV-G pseudotyped lentiviral particles that are ready to infect almost all types of mammalian cells, including primary and non-dividing cells. These viruses constitutively express human CD22 (NM_001771) under the control of an EF1A promoter (Figure 1).
Figure 1. Schematic of the lenti-vector used to generate the CD22 Lentivirus.
Two vials (500 µl x 2) of lentivirus at a titer ≥107 TU/ml. The titer will vary with each lot; the exact value will be provided with each shipment.
Formulation
The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
Background
CD22, also known as Siglec-2, is expressed on the membrane of B-cells. It acts as an inhibitory co-receptor of the B-cell receptor to control the B-cell immune response. In 2017 the FDA approved Besponsa, an antibody-drug conjugate targeting CD22, for patients with B-cell acute lymphoblastic leukemia (ALL). Additional therapies targeting CD22 are under evaluation.
Storage/Stability
Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.
Applications
Study the transient expression of human CD22 in target cells.
Generate a stable cell line expressing human CD22 with limiting dilution and puromycin selection.
Shipping Temperature
-80°C
Notes
To generate a CD22 stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve) for antibiotics selection of transduced cells.
Biosafety None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
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