Cas9 Expressing Immortalized Chicken Embryonic Fibroblast (DF1) Cell Line | T3453

abm | Cas9 Expressing Immortalized Chicken Embryonic Fibroblast (DF1) Cell Line | T3453

To generate the Cas9 Expressing Immortalized Chicken Embryonic Fibroblast (DF1) Cell Line, Cas9-2A-PuroR was cloned into pT2KCAGGS using Gibson Assembly (NEB). The DF1 cells were then transfected, and selected for using puromycin. The constitutive nuclear expression of Cas9 was validated by immunostaining using an antibody against Cas9, followed by DAPI-counterstaining. This DF1 cell line can be used to greatly streamline the process of experimental designs for future CRISPR/Cas9 knockout experiments in chick embryos.

Biosafety:

II

Organism:

Chicken

Source Organ:

Embryo

Growth Properties:

Adherent

Morphology:

Fibroblast

Clones:

N/A

Passage Number:

N/A

Population Doner:

20 - 24 hours

Seeding Density:

40,000 cells/cm²

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 medium available at abm

Preservation:

1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.
2. Storage Temperature: Liquid nitrogen vapour phase.

Quality Control:

1) Immunostaining

Tumorgenicn:

N/A

Shipping Conditions:

Dry Ice

Storage Contidions:

-180°C