An inducible CRISPR activation tool for accelerating plant regeneration

Zhang, Cuimei; Tang, Yajun; Tang, Shanjie; Chen, Lei; Li, Tong; Yuan, Haidi; Xu, Yujun; Zhou, Yangyan; Zhang, Shuaibin; Wang, Jianli; Wen, Hongyu; Jiang, Wenbo; Pang, Yongzhen; Deng, Xian; Cao, Xiaofeng; Zhou, Junhui; Song, Xianwei; Liu, Qikun

doi:10.1016/j.xplc.2024.100823

Cited by 5 publications

(3 citation statements)

References 71 publications

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“…Accordingly, ectopic expression of SlWRKY29 via CRISPR-activation has a beneficial morphogenic growth response by enhancing the formation of SE under in vitro culture conditions (most likely by activation of morphogenic genes; S3 Fig and S6 and S7 Tables). Thus, CRISPRa-edited SlWRKY29 embryogenic lines could be transformed with candidate genes involved in embryogenic competence, or explants might be transformed with sgRNAs targeting multiple promoters and co-activated together with SlWRKY29 for accelerated regeneration of edited plants [ 100 ].…”

Section: Discussionmentioning

confidence: 99%

“…Nonetheless, efficient employment of species-specific morphogenic genes encounters diverse methodological challenges (e.g. arduous processes of cloning, functional validation, co-expression of combinations of genes) [ 100 ]. In addition, when using the CaMV 35S promoter, undesired pleiotropic effects and harmful growth defects could be caused by constitutive expression of morphogenic regulators, which in turn requires to control expression of the morphogenic gene (via inducible expression, developmentally regulated expression, or by excision of the exogenous DNA) to regenerate normal-phenotype plants [ 62 ].…”

Section: Discussionmentioning

confidence: 99%

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Editing of SlWRKY29 by CRISPR-activation promotes somatic embryogenesis in Solanum lycopersicum cv. Micro-Tom

Valencia-Lozano,

Cabrera-Ponce,

Barraza

et al. 2024

PLoS ONE

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At present, the development of plants with improved traits like superior quality, high yield, or stress resistance, are highly desirable in agriculture. Accelerated crop improvement, however, must capitalize on revolutionary new plant breeding technologies, like genetically modified and gene-edited crops, to heighten food crop traits. Genome editing still faces ineffective methods for the transformation and regeneration of different plant species and must surpass the genotype dependency of the transformation process. Tomato is considered an alternative plant model system to rice and Arabidopsis, and a model organism for fleshy-fruited plants. Furthermore, tomato cultivars like Micro-Tom are excellent models for tomato research due to its short life cycle, small size, and capacity to grow at high density. Therefore, we developed an indirect somatic embryo protocol from cotyledonary tomato explants and used this to generate epigenetically edited tomato plants for the SlWRKY29 gene via CRISPR-activation (CRISPRa). We found that epigenetic reprogramming for SlWRKY29 establishes a transcriptionally permissive chromatin state, as determined by an enrichment of the H3K4me3 mark. A whole transcriptome analysis of CRISPRa-edited pro-embryogenic masses and mature somatic embryos allowed us to characterize the mechanism driving somatic embryo induction in the edited tomato cv. Micro-Tom. Furthermore, we show that enhanced embryo induction and maturation are influenced by the transcriptional effector employed during CRISPRa, as well as by the medium composition and in vitro environmental conditions such as osmotic components, plant growth regulators, and light intensity.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Editing of SlWRKY29 by CRISPR-activation promotes somatic embryogenesis in Solanum lycopersicum cv. Micro-Tom

Valencia-Lozano,

Cabrera-Ponce,

Barraza

et al. 2024

PLoS ONE

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show abstract

“…With the increasing CRISPR/Cas [clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein] toolkit, there appears to be no limit to the number of genes that can be activated via single guide RNA (sgRNA) targeting, meaning that the practical construct size limitations fade. By applying an inducible SunTag synthetic transcription activator, here containing a deadCas9 that is targeted to activate several developmental regulator gene loci by co-expressed sgRNAs, improved transformation, and regeneration was obtained in strawberry and sheepgrass ( C. Zhang et al ., 2024 ). Combined with an additional induction system, such a set-up can interrogate the effect of combinations of genes not only simultaneously but also sequentially.…”

Section: Regeneration By the Ectopic Expression Of Developmental Regu...mentioning

confidence: 99%

Appreciating animal induced pluripotent stem cells to shape plant cell reprogramming strategies

Wittmer,

Heidstra

2024

Journal of Experimental Botany

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Animals and plants have developed resilience mechanisms to effectively endure and overcome physical damage and environmental challenges throughout their life span. To sustain their vitality, both animals and plants employ mechanisms to replenish damaged cells, either directly, involving the activity of adult stem cells, or indirectly, via dedifferentiation of somatic cells that are induced to revert to a stem cell state and subsequently redifferentiate. Stem cell research has been a rapidly advancing field in animal studies for many years, driven by its promising potential in human therapeutics, including tissue regeneration and drug development. A major breakthrough was the discovery of induced pluripotent stem cells (iPSCs), which are reprogrammed from somatic cells by expressing a limited set of transcription factors. This discovery enabled the generation of an unlimited supply of cells that can be differentiated into specific cell types and tissues. Equally, a keen interest in the connection between plant stem cells and regeneration has been developed in the last decade, driven by the demand to enhance plant traits such as yield, resistance to pathogens, and the opportunities provided by CRISPR/Cas-mediated gene editing. Here we discuss how knowledge of stem cell biology benefits regeneration technology, and we speculate on the creation of a universal genotype-independent iPSC system for plants to overcome regenerative recalcitrance.

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Advancements in plant transformation: from traditional methods to cutting-edge techniques and emerging model species

Levengood,

Zhou,

Zhang

2024

Plant Cell Rep

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An inducible CRISPR activation tool for accelerating plant regeneration

Cited by 5 publications

References 71 publications

Editing of SlWRKY29 by CRISPR-activation promotes somatic embryogenesis in Solanum lycopersicum cv. Micro-Tom

Editing of SlWRKY29 by CRISPR-activation promotes somatic embryogenesis in Solanum lycopersicum cv. Micro-Tom

Appreciating animal induced pluripotent stem cells to shape plant cell reprogramming strategies

Advancements in plant transformation: from traditional methods to cutting-edge techniques and emerging model species

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