An inducible CRISPR activation tool for accelerating plant regeneration

Area of Science:

• Plant Biotechnology
• Gene Regulation
• Molecular Biology

Background:

• The CRISPR activation (CRISPR-a) system is valuable for precise gene regulation in plant research.
• Existing CRISPR-a systems may lack inducible control, limiting applications where constitutive expression is undesirable.

Purpose of the Study:

• To develop a chemically inducible CRISPR-a tool for plants, named ER-Tag.
• To assess ER-Tag's efficiency in target gene activation and multiplexed guide RNA screening.
• To investigate the impact of induced morphogenic gene activation on plant regeneration.

Main Methods:

• Combined the LexA-VP16-ER inducible system with the SunTag CRISPR-a system to create ER-Tag.
• Systematically compared various induction strategies for optimal gene activation.
• Utilized multiplexed guide RNAs for large-scale screening of plant regeneration genes.

Main Results:

• Achieved high efficiency in target gene activation using the ER-Tag system.
• Demonstrated successful screening of morphogenic genes and gene pairs involved in plant regeneration.
• Induced activation of identified genes accelerated regeneration and improved efficiency in eudicots and monocots (alfalfa, strawberry, sheepgrass).

Conclusions:

• ER-Tag provides a powerful, chemically inducible CRISPR-a tool for plant research.
• Expands the CRISPR toolset for studying gene function when inducible control is necessary.
• Offers a novel strategy to enhance plant regeneration efficiency in various species.