Fluorescent labeling of genomic loci in Drosophila imaginal discs with heterologous DNA-binding proteins.
Cell Rep Methods
; 2(3): 100175, 2022 03 28.
Article
in En
| MEDLINE
| ID: mdl-35475221
ABSTRACT
Using the Drosophila melanogaster Hox gene Ultrabithorax (Ubx) as an example, we demonstrate the use of three heterologous DNA-binding protein systems-LacI/LacO, ParB1/ParS1, and ParB2/ParS2-to label genomic loci in imaginal discs with the insertion of a small DNA tag. We compare each system, considering the impact of labeling in genomic regions (1) inside versus outside of a transcribed gene body and (2) with varying chromatin accessibility. We demonstrate the value of this system by interrogating the relationship between gene expression level and enhancer-promoter distance, as well as inter-allelic distance at the Ubx locus. We find that the distance between an essential intronic cis-regulatory element, anterobithorax (abx), and the promoter does not vary with expression level. In contrast, inter-allelic distance correlates with Ubx expression level.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Drosophila Proteins
/
Drosophila
Limits:
Animals
Language:
En
Journal:
Cell Rep Methods
Year:
2022
Document type:
Article
Affiliation country:
United States