Fluorescent labeling of genomic loci in Drosophila imaginal discs with heterologous DNA-binding proteins.

Autor:	Delker RK; Department of Biochemistry and Molecular Biophysics, Columbia University Irving Medical Center, New York, NY, USA.; Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, USA., Munce RH; Department of Biochemistry and Molecular Biophysics, Columbia University Irving Medical Center, New York, NY, USA.; Department of Genetics and Development, Columbia University Irving Medical Center, New York, NY, USA.; Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, USA., Hu M; Department of Biochemistry and Molecular Biophysics, Columbia University Irving Medical Center, New York, NY, USA., Mann RS; Department of Biochemistry and Molecular Biophysics, Columbia University Irving Medical Center, New York, NY, USA.; Department of Neuroscience, Columbia University Irving Medical Center, New York, NY, USA.; Department of Systems Biology, Columbia University Irving Medical Center, New York, NY, USA.; Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, USA.
Jazyk:	angličtina
Zdroj:	Cell reports methods [Cell Rep Methods] 2022 Mar 09; Vol. 2 (3), pp. 100175. Date of Electronic Publication: 2022 Mar 09 (Print Publication: 2022).
DOI:	10.1016/j.crmeth.2022.100175
Abstrakt:	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. Competing Interests: The authors declare no competing interests. (© 2022 The Authors.)
Databáze:	MEDLINE
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