Synthetic Regulatory Reconstitution Reveals Principles of Mammalian Cluster Regulation

Overview

Journal Science

Specialty Science

Date 2022 Jun 30

PMID 35771912

Authors

Sudarshan Pinglay

Milica Bulajic

Dylan P Rahe

Emily Huang

Ran Brosh

Nicholas E Mamrak

Benjamin R King

Sergei German

John A Cadley

Lila Rieber

Nicole Easo

Timothee Lionnet

Shaun Mahony

Matthew T Maurano

Liam J Holt

Esteban O Mazzoni

Jef D Boeke

Affiliations

Soon will be listed here.

Abstract

Precise gene expression is crucial for embryonic patterning. Intra- transcription factor binding and distal enhancer elements have emerged as the major regulatory modules controlling gene expression. However, quantifying their relative contributions has remained elusive. Here, we introduce "synthetic regulatory reconstitution," a conceptual framework for studying gene regulation, and apply it to the cluster. We synthesized and delivered variant rat clusters (130 to 170 kilobases) to an ectopic location in the mouse genome. We found that a minimal cluster recapitulated correct patterns of chromatin remodeling and transcription in response to patterning signals, whereas the addition of distal enhancers was needed for full transcriptional output. Synthetic regulatory reconstitution could provide a generalizable strategy for deciphering the regulatory logic of gene expression in complex genomes.

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