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© Research
Publication : Nature communications

Dissection of the Fgf8 regulatory landscape by in vivo CRISPR-editing reveals extensive intra- and inter-enhancer redundancy.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nature communications - 19 Jan 2021

Hörnblad A, Bastide S, Langenfeld K, Langa F, Spitz F,

Link to Pubmed [PMID] – 33469032

Link to DOI – 10.1038/s41467-020-20714-y

Nat Commun 2021 01; 12(1): 439

Developmental genes are often regulated by multiple elements with overlapping activity. Yet, in most cases, the relative function of those elements and their contribution to endogenous gene expression remain poorly characterized. An example of this phenomenon is that distinct sets of enhancers have been proposed to direct Fgf8 in the limb apical ectodermal ridge and the midbrain-hindbrain boundary. Using in vivo CRISPR/Cas9 genome engineering, we functionally dissect this complex regulatory ensemble and demonstrate two distinct regulatory logics. In the apical ectodermal ridge, the control of Fgf8 expression appears distributed between different enhancers. In contrast, we find that in the midbrain-hindbrain boundary, one of the three active enhancers is essential while the other two are dispensable. We further dissect the essential midbrain-hindbrain boundary enhancer to reveal that it is also composed by a mixture of essential and dispensable modules. Cross-species transgenic analysis of this enhancer suggests that its composition may have changed in the vertebrate lineage.

https://pubmed.ncbi.nlm.nih.gov/33469032