CRISPR Activator Approaches to Study Endogenous Androglobin Gene Regulation

Koay TW, Schödel J, Hoogewijs D (2023)

Publication Type: Book chapter / Article in edited volumes

Publication year: 2023


Publisher: Humana Press Inc.

Series: Methods in Molecular Biology

Book Volume: 2648

Pages Range: 167-185

DOI: 10.1007/978-1-0716-3080-8_11


Androglobin (ADGB), the most recently identified member of the mammalian globin family, is a chimeric protein with an unusual, embedded globin domain that is circularly permutated and exhibits hallmarks of a hexacoordinated heme-binding scheme. Whereas abundant expression of ADGB was initially found to be mainly restricted to cells in the postmeiotic stages of spermatogenesis, more recent RNA-Seq-based expression analysis data revealed that ADGB is detectable in cells carrying motile cilia or flagella. This very tight regulation of ADGB gene expression urges the need for alternative techniques to study endogenous expression in classical mammalian cell models, which do not express ADGB. We describe here the use of CRISPR activation (CRISPRa) technology to induce endogenous ADGB gene expression in HEK293T, MCF-7, and HeLa cells from its promoter and illustrate how this method can be employed to validate putative regulatory DNA elements of ADGB in promoter and enhancer regions.

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Koay, T.W., Schödel, J., & Hoogewijs, D. (2023). CRISPR Activator Approaches to Study Endogenous Androglobin Gene Regulation. In (pp. 167-185). Humana Press Inc..


Koay, Teng Wei, Johannes Schödel, and David Hoogewijs. "CRISPR Activator Approaches to Study Endogenous Androglobin Gene Regulation." Humana Press Inc., 2023. 167-185.

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