Zbed6 Knockout C2C12 Cell Line

The Zbed6 Knockout C2C12 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the C2C12 mouse myoblast background, featuring targeted disruption of the Zbed6 gene. This loss-of-function model enables investigation of Zbed6-mediated transcriptional regulation in muscle biology. The knockout results in disrupted Zbed6 expression and provides a stable tool for studying Zbed6 depletion in myogenic cells.

The parental C2C12 cell line is a well-established subclone of the C2 mouse myoblast line, derived from satellite cells of C3H mouse muscle. These myoblasts proliferate under high-serum conditions and differentiate into multinucleated myotubes upon serum withdrawal, serving as an in vitro model for skeletal muscle differentiation. The C2C12 line retains muscle-specific protein expression and recapitulates key myogenic steps, making it ideal for studying muscle development mechanisms.

Zbed6 encodes a zinc finger BED domain transcription factor that represses Igf2 expression. In C2C12 cells, Zbed6 binds the Igf2 promoter, suppressing its transcription and limiting Igf2 availability. Igf2 activates the Igf1r/Akt/mTOR pathway, promoting myoblast proliferation and differentiation via MyoD and Myogenin. Thus, Zbed6 is a critical brake on Igf2-Igf1r-Akt-mTOR signaling, modulating the balance between myoblast maintenance and differentiation. The Zbed6-Igf2 regulatory interaction is a key node in the myogenic network, potentially influenced by upstream myogenic factors like MyoD.

Disruption of Zbed6 in C2C12 cells derepresses Igf2, increasing Igf2 protein and enhancing Igf1r/Akt/mTOR signaling. This leads to increased activation of myogenic transcription factors and accelerated myotube formation. The Zbed6 knockout C2C12 line thus serves as a tool to dissect the inhibitory regulation of the Igf2 axis and its interface with signals controlling muscle fiber size and regeneration. By removing the endogenous repressor, researchers can assess the impact of unchecked Igf2 activity on myoblast behavior and myotube hypertrophy in vitro.

This cell line is suited for myotube differentiation assays, immunoblotting for Igf2 and myogenic markers, RT-qPCR for Zbed6 and Igf2, MyHC immunofluorescence, and Igf2 promoter reporter assays. It provides a platform for screening muscle growth modulators, evaluating therapies for muscle wasting, and investigating transcriptional control of Igf2 in hypertrophy and development. For further information, please contact Ascent Research.