CRISPR Activator Approaches to Study Endogenous Androglobin Gene Regulation

Koay, Teng Wei; Schödel, Johannes; Hoogewijs, David

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

Cited by 2 publications

(2 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Arthropods display a heavily rearranged or degenerated globin domain inside the chimeric Adgb protein 9,31 . Adgb is detectable in cells carrying motile cilia or flagella and seems to be essential in spermatogenesis and ciliogenesis 29,40,41 . In contrast to the vertebrate‐specific Gbs, which are well documented and whose basic functions are well known, those of Adgb, Ngb, and GbX are still poorly understood.…”

Section: Resultsmentioning

confidence: 99%

Globin phylogeny, evolution and function, the newest update

Schuster,

Salvatore,

Moens

et al. 2024

Proteins

View full text Add to dashboard Cite

Our globin census update allows us to refine our vision of globin origin, evolution, and structure to function relationship in the context of the currently accepted tree of life. The modern globin domain originates as a single domain, three‐over‐three α‐helical folded structure before the diversification of the kingdoms of life (Bacteria, Archaea, Eukarya). Together with the diversification of prokaryotes, three monophyletic globin families (M, S, and T) emerged, most likely in Proteobacteria and Actinobacteria, displaying specific sequence and structural features, and spread by vertical and horizontal gene transfer, most probably already present in the last universal common ancestor (LUCA). Non‐globin domains were added, and eventually lost again, creating multi‐domain structures in key branches of M‐ (FHb and Adgb) and the vast majority of S globins, which with their coevolved multi‐domain architectures, have predominantly “sensor” functions. Single domain T‐family globins diverged into four major groups and most likely display functions related to reactive nitrogen and oxygen species (RNOS) chemistry, as well as oxygen storage/transport which drives the evolution of its major branches with their characteristic key distal residues (B10, E11, E7, and G8). M‐family evolution also lead to distinctive major types (FHb and Fgb, Ngb, Adgb, GbX vertebrate Gbs), and shows the shift from high oxygen affinity controlled by TyrB10‐Gln/AsnE11 likely related to RNOS chemistry in microorganisms, to a moderate oxygen affinity storage/transport function controlled by hydrophobic B10/E11–HisE7 in multicellular animals.

show abstract

Section: Resultsmentioning

confidence: 99%

Globin phylogeny, evolution and function, the newest update

Schuster,

Salvatore,

Moens

et al. 2024

Proteins

View full text Add to dashboard Cite

show abstract

“…Candidate gRNAs targeting the first 100 bps upstream of the ADGB promoter [16], as well as control gRNAs targeting more distal regions, were used to interfere with the binding of MYBL2. For this purpose, 750 ng pLenti6.2/V5-DEST-MYBL2 were co-transfected with 1000 ng pcDNA3-dCas9, 600 ng gRNA, 500 ng of the ADGB promoter fragment pGL3-AP431 (and 25 ng of pRL-SV40 Renilla into HEK293T cells (12-well plates).…”

Section: Dcas9-mediated Interference Of Transcription Factor Bindingmentioning

confidence: 99%

Ectopic MYBL2-Mediated Regulation of Androglobin Gene Expression

Herwig,

Osterhof,

Keppner

et al. 2024

Cells

Self Cite

View full text Add to dashboard Cite

Androglobin (ADGB) is a highly conserved and recently identified member of the globin superfamily. Although previous studies revealed a link to ciliogenesis and an involvement in murine spermatogenesis, its physiological function remains mostly unknown. Apart from FOXJ1-dependent regulation, the transcriptional landscape of the ADGB gene remains unexplored. We, therefore, aimed to obtain further insights into regulatory mechanisms governing ADGB expression. To this end, changes in ADGB promoter activity were examined using luciferase reporter gene assays in the presence of a set of more than 475 different exogenous transcription factors. MYBL2 and PITX2 resulted in the most pronounced increase in ADGB promoter-dependent luciferase activity. Subsequent truncation strategies of the ADGB promoter fragment narrowed down the potential MYBL2 and PITX2 binding sites within the proximal ADGB promoter. Furthermore, MYBL2 binding sites on the ADGB promoter were further validated via a guide RNA-mediated interference strategy using reporter assays. Chromatin immunoprecipitation (ChIP)-qPCR experiments illustrated enrichment of the endogenous ADGB promoter region upon MYBL2 and PITX2 overexpression. Consistently, ectopic MYBL2 expression induced endogenous ADGB mRNA levels. Collectively, our data indicate that ADGB is strongly regulated at the transcriptional level and might have functions beyond ciliogenesis.

show abstract

CRISPR Activator Approaches to Study Endogenous Androglobin Gene Regulation

Cited by 2 publications

References 21 publications

Globin phylogeny, evolution and function, the newest update

Globin phylogeny, evolution and function, the newest update

Ectopic MYBL2-Mediated Regulation of Androglobin Gene Expression

Contact Info

Product

Resources

About