Endogenous tagging of the TBXT (Brachyury) gene with 2A-EGFP in BJNhem19 human embryonic stem cells using CRISPR-Cas9 genome editing.

CRISPR/Cas9 editing created a Brachyury/TBXT-2A-EGFP reporter line in human embryonic stem cells, enabling real-time monitoring of mesodermal differentiation.

• Why it matters: Understanding mesodermal lineage specification is crucial for developmental biology and regenerative medicine, but tools for live tracking of this process are limited.
• What they did: The authors used CRISPR/Cas9-mediated homology-directed repair to insert the EGFP reporter into the TBXT gene in BJNhem19 hESCs, confirming integration by DNA sequencing and expression comparison.
• The result: This reporter line offers a fluorescence-based platform for real-time monitoring of mesodermal commitment, facilitating high-content screening of differentiation protocols.