ExperimentMic multiplexed FISH on 293T

multiplexed FISH4DNEXZ1SRCPW
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Multiplexed FISH in HEK293T reporter cell line treated with doxycyline for 5 days, causing recruitment of KRAB mutant (EEW25AAA) which unlike wt-KRAB is unable to silence reporter gene expression, used as negative control, Biological Replicate 2 ,Technical Replicate 1 (control and experiment relationships are described in detailed in 4DNFIWQ9QBOB supplementary file).

   February 4th, 2025 at 9:44pm
Source Publication
Single-cell chromatin state transitions during epigenetic memory formation.
Fujimori T, Rios-Martinez C, et al., bioRxiv : the preprint server for biology 2023

Properties  

Experiment Type 
multiplexed FISH
Follows SOP 
No
Biosample 
4DNBSGAUU15D
Biosample Biosource 
293T - 4DNSRXG828T6
Biosample Modifications 
Viral transduction insertion of reporter construct integrated into AAVS1 safe harbor site on chr19 and Viral transduction insertion of mutant (EEW25AAA) KRAB domain fused with rTetR
Biosample Treatments 
doxycycline treatment (1.0 μg/mL, 5.0d)
Microscopy Technique 
Widefield
Microscope Quality Control 
None
Microscope Configuration 
None
Channel
Imaging Rounds
Path
 
ch00
-
DAPI staining
ch01
1-19
Custom genome assembly (mCitrine reporter + 9 TetO sites inserted into AAVS1 safe harbor site 55115766 on chr19) targeted by Cy5-labeled imaging oligos
ch02
1-19
Custom genome assembly (mCitrine reporter + 9 TetO sites inserted into AAVS1 safe harbor site 55115766 on chr19) targeted by Cy3-labeled fiducial oligos

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