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HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries

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Published on: March 31, 2019

Functional analysis of CTCF during mammalian limb development.

Natalia Soshnikova¹, Thomas Montavon, Marion Leleu

¹Department of Zoology and Animal Biology, University of Geneva, Switzerland.

Developmental Cell

|December 15, 2010

Summary

CCCTC-binding factor (CTCF) is crucial for cell survival during limb development, not primarily as an insulator. Its depletion causes apoptosis and limb loss, highlighting its role in cellular homeostasis.

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Area of Science:

Molecular Biology
Developmental Biology
Genetics

Background:

CCCTC-binding factor (CTCF) is a nuclear protein known for its role as a transcriptional insulator.
CTCF-binding sites are implicated in isolating gene clusters, such as Hox genes, from regulatory interference.
Proper regulation of Hox gene clusters is essential during embryonic development, particularly in limb formation.

Purpose of the Study:

To investigate the role of CTCF in limb development, specifically its proposed insulating function on Hox gene clusters.
To determine the necessity of CTCF for maintaining Hox gene expression patterns during limb formation.
To elucidate the broader function of CTCF in the context of limb development beyond transcriptional insulation.

Main Methods:

Conditional inactivation of the Ctcf gene in mouse incipient forelimbs.
Analysis of Hoxd gene expression patterns following Ctcf deletion.
Genome-wide transcriptome analysis using tiling arrays to assess global gene regulation changes.
Histological examination to evaluate limb structure and cell viability.

Main Results:

Conditional Ctcf inactivation did not alter the overall Hoxd gene expression pattern during limb development.
Transcriptome analysis revealed minimal impact of CTCF depletion on global gene regulation.
Ctcf deletion led to massive apoptosis and severe limb malformation, indicating a critical role in cell survival.
CTCF is essential for maintaining cellular homeostasis through the regulation of critical target genes.

Conclusions:

CTCF's primary function in this physiological context is not as a transcriptional insulator for Hox genes.
CTCF is indispensable for cell survival during limb development.
CTCF regulates genes critical for cellular homeostasis, and its absence leads to developmental failure.