CRISPR Crops: Plant Genome Editing Toward Disease Resistance

Abstract

Genome editing by sequence-specific nucleases (SSNs) has revolutionized biology by enabling targeted modifications of genomes. Although routine plant genome editing emerged only a few years ago, we are already witnessing the first applications to improve disease resistance. In particular, CRISPR-Cas9 has democratized the use of genome editing in plants thanks to the ease and robustness of this method. Here, we review the recent developments in plant genome editing and its application to enhancing disease resistance against plant pathogens. In the future, bioedited disease resistant crops will become a standard tool in plant breeding.

In planta gene targeting can be enhanced by the use of CRISPR/Cas12a

“…The emergence of the CRISPR/Cas technology enabled targeted double‐strand break (DSB) induction at high efficiency in a simple way (Jinek et al ., ; Le Cong et al ., ). Since then, a broad range of new applications of the technology is currently transforming plant biology (Baltes et al ., ; Malzahn et al ., ; Puchta, ; Kumlehn et al ., ; Langner et al ., ; Chen et al ., ; Sedeek et al ., ; Wolter et al ., ). The stimulatory potential of targeted DSB induction for non‐homologous end‐joining (NHEJ)‐mediated gene knockout and homologous recombination (HR)‐mediated gene targeting (GT) in plants was already demonstrated long before the development of CRISPR/Cas (Puchta et al ., ; Salomon and Puchta, ).…”

Multiplexed CRISPR-Cas9 based genome editing ofRhodosporidium toruloides

“…However, deletion of CAR2 was markedly more successful in even the worst-case scenario (3.4% ± 2.7%), while the best-case scenario resulted in roughly a one-to-one ratio of white to red colonies. The locus-dependency of Cas9 editing efficiency has been noted in other non-yeast eukaryotic systems (23, 59, 60). Combined with the qualitatively large differences observed between editing efficiencies of Cas9 at various URA3 target locations, optimization of the Cas9 target sequence appears to be particularly important for editing in R. toruloides.…”

“…Concurrent with these advances is a revolution in the field of genome engineering brought on by the development of precise genome and transcriptome editing through clustered regularly interspaced short palindromic repeat (CRISPR)-based systems (20–22). Modern genetic engineering approaches in other organisms have largely gravitated towards using CRISPR systems to enact desired DNA changes (23–25). This approach utilizes a ribonucleoprotein complex (RNP) consisting of a CRISPR associated nuclease (Cas, the most common being Cas9) and a synthetic single guide RNA (sgRNA).…”

Metabolomics should be deployed in the identification and characterization of gene‐edited crops

“…The most recent technology is based on a form of bacterial adaptive immunity, in which previously encountered invasive DNA sequences are committed to molecular memory and targeted in future challenges by expressing CRISPRs representing DNA fragments captured from invading pathogens. The resultant CRISPR RNAs act as guides for CRISPR‐associated (Cas) nucleases that attack the pathogens upon subsequent infection (Knott and Doudna, ; Langner et al ., ). Researchers have exploited this process by constructing synthetic guide RNAs that direct the Cas nuclease to genomic targets.…”

“…The last decade has been characterized by the adoption of genome‐editing systems following the revolutionary discovery of transcriptional activator‐like effector (TALE) proteins, which are more suitable for the precise engineering of targeted DNA sequences (Stella and Montoya, ) and the subsequent widespread adoption of the clustered regularly interspaced short palindromic repeats (CRISPR)‐associated protein (Cas) system (Puchta and Fauser, ; Scheben et al ., ; Yin et al ., ). We refer the reader to the plethora of excellent reviews on plant gene editing (Kim and Kim, ; Baltes and Voytas, ; Bortesi et al ., ; Wright et al ., ; Pacher and Puchta, ; Zhu et al ., ; Knott and Doudna, ; Langner et al ., ) for details of the underlying molecular mechanisms and the manifold applications. Here we outline the opportunities that these techniques afford as well as review recent additions to the repertoire of applications.…”