ABSTRACT
The transcription factor HAND1 is a critical regulator of cardiac development which is expressed in sub-populations of cardiac progenitors and cardiomyocytes. The transcription factor NKX2-5, in contrast, is expressed more widely in cardiac cells. Here we report the generation of a dual reporter hESC line where the expression of these genes can be simultaneously measured, enabling lineage analysis as well as studies of HAND1 and NKX2-5 gene regulation and protein function. This tool will have wide utility particularly for research on developmental biology and disease modelling.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors , Homeobox Protein Nkx-2.5 , Homeodomain Proteins , Human Embryonic Stem Cells , Transcription Factors , Humans , Homeobox Protein Nkx-2.5/metabolism , Homeobox Protein Nkx-2.5/genetics , Transcription Factors/metabolism , Transcription Factors/genetics , Human Embryonic Stem Cells/metabolism , Human Embryonic Stem Cells/cytology , Homeodomain Proteins/metabolism , Homeodomain Proteins/genetics , Basic Helix-Loop-Helix Transcription Factors/metabolism , Basic Helix-Loop-Helix Transcription Factors/genetics , Cell Line , Green Fluorescent Proteins/metabolism , Green Fluorescent Proteins/genetics , Myocytes, Cardiac/metabolism , Myocytes, Cardiac/cytology , Genes, Reporter , Cell Differentiation , Cell LineageABSTRACT
The transcription factor NKX2-5 is a highly conserved master regulator of heart development which is widely expressed in cardiac progenitors and cardiomyocytes. Fluorescent reporters of NKX2-5 that minimally perturb normal protein expression can enable the identification, quantification and isolation of NKX2-5-expressing cells in a normal physiological state. Here we report the generation of two new hESC lines with eGFP inserted upstream (5') or downstream (3') of NKX2-5, linked by a cleavable T2A peptide. These complementary reporters produce a robust fluorescent signal in cardiac cells and have wide utility particularly for research on developmental biology and disease modelling.