RPE1 FRT/TR Cell Line

This cell line can be used to generate cell lines with tetracycline inducible expression of genes of interest. The cell line has a randomly integrated FRT locus and Tet repressor for Flp-In expression. For example the originator of the cell line used this cell line to generate tetracycline-inducible cell lines expressing APC15-IRES2-mRuby, APC15-3xflag and Cyclin […]

Contributors

Inventor Institute
Jonathon Pines University of Cambridge
Cat. #: 153242
Unit size: 1×10^6 cells / vial
Research Fields: Genetics
Organism: Human
Tissue: Eye
Growth properties: Adherent cell line
Product description: This cell line can be used to generate cell lines with tetracycline inducible expression of genes of interest. The cell line has a randomly integrated FRT locus and Tet repressor for Flp-In expression. For example the originator of the cell line used this cell line to generate tetracycline-inducible cell lines expressing APC15-IRES2-mRuby, APC15-3xflag and Cyclin B1-L45A-HA using the FLIP-in system and a modified pCDNA5/FRT/TO vector. This cell line is resistant to Zeocin (from FRT) and Blasticidin (from TR). It also carries resistance to hygromycin and puromycin, inherited from the parental RPE1 cell line. After Flp-In integration, selection is performed using Geneticin (a neomycin analogue), because the hygromycin resistance cassette in the original pcDNA5/FRT/TO plasmid was replaced with a neomycin resistance marker. The FRT and TR cassettes are stably integrated, so additional co-selection with Blasticidin or Zeocin is not required.
Conditional: Yes
Conditional description: Tetracycline-inducible expression of genes of interest.
Production details: This tetracycline-inducible RPE1 cell line was created by random integration of an FRT site and a Tet repressor gene into retinal pigment epithelial 1 (RPE1) cells. The inventors used electroporation for transfection, specifically with Invitrogen’s Neon transfection system. Cells were allowed to recover for 2 –3 days post-transfection, then split into media containing Geneticin at a final concentration of 0.5 mg/mL (ThermoFisher Scientific, 910131027). This selection medium was refreshed every 2–3 days for approximately 10–14 days until cell death ceases. Standard media included high-quality, tetracycline-free FBS. For plasmid ratios during transfection, a 4:1 ratio of pOG44 (Flp-recombinase expression plasmid) to the gene-of-interest plasmid (pcDNA5/FRT/TO/Neo + gene of interest) was used.
Cellosaurus ID: CVCL_VP32
Parental cell line: RPE1
Format: Frozen
Storage conditions: Liquid Nitrogen
Shipping conditions: Dry ice
Growth medium: 500 mL Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham (Sigma: D6241), 17.3 mL of 7.5% sodium bicarbonate, 10% FBS, 1% Pen/Strep, 1% 200 mM Glutamax and 1 mL Amphotericin B.
Subculture routine: Please see detailed protocol within the Product Datasheet in the Documentation section below.
Mycoplasma free: Yes
Biosafety level: 1
References:

Mansfield et al. 2011. Nat Cell Biol. 13:1234-43. PMID: 21926987

Related Tools for Cell lines

Cat. # Tool Name
151445 1-7HB2 Cell Line Key Info View Tool
151446 FLYRD18 Cell Line Key Info View Tool
151558 MEF Trex1 KO Cell Line Key Info View Tool
151630 PDVC57B Cell Line Key Info View Tool
151663 MEF PKCe KO KI Cell Line Key Info View Tool

Related News

UM-UC-10 Cell Line. Image courtesy of the European Collection of Authenticated Cell Cultures (ECACC), UK.
Cell Lines

Popular cell lines now available only from CancerTools.org

Read More
Cell Lines

New cell lines: murine melanocyte and melanoblast cell lines from Dr. Elena Sviderskaya

Read More
Image Reference: 2x PTEN -/- mouse prostate cancer cell lines (MuCap system). Histology staining of syngraft tumours (Van Duijn et al., 2018).
Cell Lines

38 new prostate cancer cell lines

Read More

Tool enquiry

Please ensure you use your organisation email address rather than personal where possible, as this helps us locate your organisation in our system faster.

Please note we may take up to three days to respond to your enquiry.

CancerTools.org uses the contact information provided to respond to you about our research tools and service. For more information please review our privacy policy.