Labeling the TBXT gene with a glowing marker in human embryonic stem cells using CRISPR gene editing

A Brachyury/TBXT-2A-EGFP knock-in reporter line was successfully generated in human embryonic stem cells.

• Brachyury (TBXT) is crucial for guiding human embryonic stem cells toward mesodermal lineage specification.
• The reporter line was created using CRISPR/Cas9-mediated homology-directed repair.
• Integration of the reporter cassette was confirmed by DNA sequencing.
• EGFP expression was compared to endogenous TBXT gene expression during mesodermal differentiation.
• This system allows for real-time monitoring of mesodermal commitment.

Simplified