Inducible CRISPR Activation Tool To Promote Plant Regeneration

On January 18, 2024, Plant Communications published an article titled "An Inducible CRISPR-activation tool for accelerated plant regeneration" online. This study achieved an improvement in the genetic transformation efficiency and regeneration efficiency of different monocotyledonous and dicotyledonous plants through precise activation of plant endogenous tissue regeneration-related genes.

In recent years, with the rapid development of gene editing technology, establishing an efficient and broad-spectrum plant regeneration system is of great significance to promote plant breeding and improvement. However, limited by species and genotypes, the regeneration efficiency of most plants is relatively low. Previous studies have shown that ectopic expression of regeneration-related genes in somatic cells can significantly improve the regeneration efficiency of a variety of difficult-to-regenerate crops. Nonetheless, overexpression of some regeneration-related transcription factors may lead to phenotypic abnormalities in transgenic plants. At the same time, given the large number of members of the pro-regeneration transcription factor family, traditional screening of regeneration-related genes in specific species requires tedious steps, including cloning, functional verification, and co-expression of morphogenesis gene combinations. Therefore, there is an urgent need to develop some new tools to overcome these technical difficulties and further improve plant regeneration efficiency.

CRISPR/dCas9 is a genetic manipulation tool derived from CRISPR/Cas9. The CRISPR/dCas9 system mainly achieves the regulation of gene expression through the CRISPR activation (CRISPRa) or CRISPR interference (CRISPRi) system. Among them, the CRISPRa system is a powerful tool for transcriptional activation of endogenous genes. In addition, the β-estradiol-inducible XVE (LexA-VP16-ER) system is a gene expression regulation system widely used in plants. Traditionally, XVE systems can typically regulate only one gene at a time, making large-scale gene function testing a challenge. By combining it with CRISPR activation tools, precise activation of multiple endogenous genes can be achieved.

Based on this, this study developed a set of phytochemically induced CRISPRa tools, called “ER-Tag”, by combining the XVE induction system with the SunTag CRISPRa system. This study systematically compared the efficiency of coupling strategies between different XVE systems and the SunTag CRISPRa system to activate regeneration-related genes. This study realizes the feasibility of large-scale screening of morphogenetic genes and gene combinations related to Arabidopsis regeneration under "sub-optimal hormone" culture conditions. In addition, this study screened the optimal combination of morphogenetic genes to improve the regeneration efficiency of woodland strawberry and sheepgrass by co-activating randomly paired morphogenetic genes, thus proving that the “ER-Tag” strategy can be applied to improve the regeneration efficiency of dicotyledonous and monocotyledonous plants. This study expands the application scope of plant CRISPR tools. This tool is not only limited to applications in the field of plant regeneration, but may also be used to precisely regulate genes in other biological processes and promote more in-depth research on gene functions.

Agrobacterium Competent Cells We Offer

Reference:

Zhang, C., et al. An inducible CRISPR activation tool for accelerating plant regeneration. Plant Commun. 2024 Jan 18: 100823.

Related Services & Products

• Plant Breeding
• Plant Genetic Engineering
• Plant Genetic Transformation
• Agrobacterium Competent Cells
• Gene Overexpression in Plants
• CRISPR Activation (CRISPRa) Service
• CRISPR Interference (CRISPRi) Service