CRISPR screens in plants: approaches, guidelines, and future prospects

Area of Science:

• Plant Biology
• Genomics
• Genome Engineering

Background:

• CRISPR-associated systems have transformed genome engineering and functional genomics.
• CRISPR screens allow for high-throughput gene function testing at a genomic scale.
• Implementation of CRISPR screens in plants is a developing field with significant potential.

Purpose of the Study:

• To discuss concepts, tools, and workflows for establishing CRISPR screens in plants.
• To analyze recent reports on CRISPR screens for generating mutant collections and diversifying DNA sequences.
• To provide insights into designing CRISPR knockout screens in plants, considering challenges and limitations.

Main Methods:

• Review of general concepts, tools, and workflows for CRISPR screens in plants.
• Analysis of existing literature on CRISPR-based mutant collections and DNA diversification.
• Examination of design options for CRISPR knockout screens, including multiplexing strategies.

Main Results:

• CRISPR screens offer powerful new methods for interrogating gene regulatory networks in plants.
• Recent reports demonstrate the use of CRISPR screens for generating mutant collections and diversifying DNA sequences.
• The multiplexing capabilities of CRISPR screens are particularly valuable for studying gene redundancy in complex plant genomes.

Conclusions:

• CRISPR screens are poised to significantly advance plant functional and synthetic biology.
• The integration of CRISPR screens with genomic data will enable deeper analysis of plant genomes.
• Combinatorial mutant screens can routinely generate higher-order mutant collections for gene network characterization.