Stable ΔN-IκBα Expressing HOS-G (HOS-G-ΔN-IκBα) Cell Line | T6108

abm | Stable ΔN-IκBα Expressing HOS-G (HOS-G-ΔN-IκBα) Cell Line | T6108

Human Osetosarcoma (HOS) cells were transfected with a Lentivirus vector to stably express a degradation resistant I-κBα (∆N-I-κBα) under the control of a SV40 promoter. Successfully transfected cells were selected for puromycin resistance. As a result of the constitutive repression of NF-κB by the degradation resistant inhibitor ∆N-I-κBα, these cells are resistant to senescence caused by infection with human T-lymphotropic virus type 1 (HTLV-1). HOS-G/ΔN-IκBα line can be productively infected by HTLV-1 and can spread HTLV-1 to susceptible cells. This line also contains a reporter cassette consisting of 18 copies of Tax-inducible HTLV-1 21bp repeat, the HTLV-1 TATA element, the complete R region, and part of the U5 sequence fused with enhanced green fluorescence protein (eGFP). As a result of this reporter element the expression of eGFP is a positive indicator of successful HTLV-1 infection in cells. It is suggested that this cell line can be used to study of HTLV-1 replication in cell culture.

Biosafety:

II

Organism:

Human

Source Organ:

Bone

Growth Properties:

Adherent

Morphology:

Bipolar

Clones:

N/A

Passage Number:

N/A

Population Doner:

N/A

Seeding Density:

N/A

Markers:

puromycin

Applications:

For Research Use Only

Doner Gender:

N/A

Donor Ethnicity:

N/A

Knockdown Method:

N/A

Induction:

N/A

Overexpression:

N/A

Freeze Thaw:

N/A

Propagation:

The base medium for this cell line is Prigrow III available at abm

Preservation:

N/A

Quality Control:

1) Verified the expression of GFP upon HTLV-1 infection via imaging (Figure 1). 2) Verified the expression of ΔN-IκBα in Western blot (Figure 1). 3) Verified that cell line continues to propagate after HTLV-1 infection (Figure 2). 4) Confirmed the expression of viral genes Tax, Rex and Gag through Western Blot analysis ( HTLV-1 proteins) (Figure 3A). 5) Verified viral spread to B-cells through Electron microscopy (Figure 3B). 6) Showed cells could spread from one HOS line to another as well as to T-cell reporter line by imaging RFP expression and measuring Luciferase activity of the T-cell reporter line (Figure 5).

Tumorgenicn:

N/A

Shipping Conditions:

Dry Ice

Storage Contidions:

-180°C