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Xist-TetOP mutant: ΔB cell line

Invented by Simão Teixeira da Rocha
Invented at Instituto de Medicina Molecular João Lobo Antunes
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Figure 1 – Xist-TetOP ΔB XY ESCs (source: Bousard et al., 2019 PMID: 31356285). A. Deletion mapping by Sanger sequencing and expression analysis across the deleted region; the scheme represents only the first exon of Xist, red arrows indicate forward primers, and green arrows represent reverse primers; the primer on the left is the sequencing primer for each mutant; B means PCR blank. B. Representative image of combined immunofluorescence (IF for H3K27me3 or H2AK119ub (green) with RNA FISH for Xist (red) in Xist ΔB mutant at day 2 of differentiation in DOX conditions; blue - DAPI staining
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Image thumbnail for Xist-TetOP mutant: ΔB cell line
Catalogue Number 160726
Antigen/Gene or Protein Targets Mouse Xist lncRNA
Parental Line Xist-TetOP XY ESCs (Wutz et al., 2002 – PMID: 11780141)
Host Mouse
Model Stem Cells
Relevance Deletion of mouse Xist lncRNA to dissect specific RNA modules in the process of X-chromosome inactivation.
Production Details CRISPR/Cas9 genome editing of the Xist-TetOP XY ESC line
Conditional No
Conditional Description Inducible through addition of doxycycline (1.5 μg/ml) to the medium
Research Area Epigenetics & Nuclear Signalling, Stem Cell Biology
Growth/Phenotype Keywords Partial disruption of Polycomb recruitment to the inactive X-chromosome.
Recommended Growing Conditions ESCs grow in feeder-free classic ESC medium - DMEM media containing 15% fetal bovine serum (FBS), 103 U/ml leukemia inhibitor factor (LIF), 10−4 mM 2‐mercaptoethanol, 50 U/ml penicillin, and 50 μg/ml of streptomycin (Gibco) at 37°C in 5-8% CO2. Medium is changed daily Inducible expression of Xist driven by a TetO promoter is achieved by adding DOX (1.5 μg/ml).
Positive Control Yes, Xist-TetOP XY ESCs (Wutz et al., 2002 – PMID: 11780141)
Notes Cancer Research Technology Limited (trading research tools as Ximbio) has been granted a non-exclusive license to the CRISPR-Cas9 technology by ERS Genomics Ltd under the patent rights listed here.

This license from ERS Genomics Ltd allows Ximbio to develop and commercialise CRISPR-Cas9 modified cell lines for research use only. Ximbio can provide these modified CRISPR-Cas9 cell lines to companies under a label-use only license.

References

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References: 1 entry

Bousard et al. 2019. EMBO Rep. 20(10):e48019. PMID: 31456285.

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References: 1 entry

Bousard et al. 2019. EMBO Rep. 20(10):e48019. PMID: 31456285.

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