Plant Functional Genomics Based on High‐Throughput CRISPR Library Knockout Screening: A Perspective

Abstract

Plant biology studies in the post‐genome era have been focused on annotating genome sequences’ functions. The established plant mutant collections have greatly accelerated functional genomics research in the past few decades. However, most plant genome sequences' roles and the underlying regulatory networks remain substantially unknown. Clustered, regularly interspaced short palindromic repeat (CRISPR)‐associated systems are robust, versatile tools for manipulating plant genomes with various targeted DNA perturbations, providing an excellent opportunity for high‐throughput interrogation of DNA elements’ roles. This study compares methods frequently used for plant functional genomics and then discusses different DNA multi‐targeted strategies to overcome gene redundancy using the CRISPR‐Cas9 system. Next, this work summarizes recent reports using CRISPR libraries for high‐throughput gene knockout and function discoveries in plants. Finally, this work envisions the future perspective of optimizing and leveraging CRISPR library screening in plant genomes' other uncharacterized DNA sequences.

Advances in genome sequencing and artificially induced mutation provides new avenues for cotton breeding

“…(2024) used CRISPR/Cas9 and WGS to create and analyze insect-resistant cotton mutants, elucidating molecular pathways contributing to resistance. He et al. (2023) highlighted the potential of CRISPR-based methods for large-scale gene knockout screening and uncovering gene functions and regulatory networks.…”