SHIELD: a platform for high-throughput screening of barrier-type DNA elements in human cells

Area of Science:

• Genomics
• Epigenetics
• Molecular Biology

Background:

• Chromatin boundary elements define genome topology and regulate gene expression.
• DNA insulators, derived from boundary elements, are crucial for stable transgene expression.
• Characterizing non-coding boundary elements is challenging due to a lack of screening platforms.

Purpose of the Study:

• To develop a high-throughput platform for screening barrier-type DNA elements.
• To discover novel DNA elements with insulator properties from the human non-coding genome.

Main Methods:

• Developed SHIELD (Site-specific Heterochromatin Insertion of Elements at Lamina-associated Domains) for screening.
• Utilized serine integrase specificity for heterochromatin insertion within lamina-associated domains (LADs).
• Screened 1000 DNA elements for barrier activity in human cells.

Main Results:

• Identified 8 novel DNA elements with significant barrier activities.
• The identified elements showed comparable activity to the established cHS4 insulator core region.
• Demonstrated the utility of SHIELD for high-throughput evaluation of DNA barrier elements.

Conclusions:

• SHIELD is an effective platform for discovering potent barrier DNA elements.
• Novel barrier elements can be identified from the non-coding genome using SHIELD.
• This platform facilitates the search for new insulators for stable gene expression applications.