Overall, the editing efficiency of gRNAs observed in our study correlated with predictive scores from the gRNA design tools. Misregulation of the chromatin remodelers following the heat shock treatment could potentially be involved in the increase of editing efficiency in wheat. We also examined the expression of DNA repair and replication gene orthologues in response to heat shock in wheat leaves. Only one out of three gRNAs demonstrated significantly increased editing efficiency following heat shock treatment.

We selected three guide RNAs (gRNAs) and evaluated the potential of using heat shock at the seedlings stage to increase editing efficiency in transgenic plants. Here we examined factors that affect the CRISPR/Cas9-mediated gene editing efficiency in allohexaploid wheat plants.

Gene editing in polyploid crops still suffers from low efficiency, and further improvement is needed for its routine implementation in the modern breeding practice. This review highlights current advances in CRISPR/Cas strategies employed in the enhancement of plant yield and examines their contribution to the development of more sustainable agricultural production. The development of plants that are resistant or tolerant to biotic or abiotic stress is complex, and the CRISPR/Cas system can offer an alternative way to develop plants resistant to abiotic and biotic stresses. In general, plant development can be negatively influenced by different factors, such as biotic and abiotic stress. This system can be considered a robust gene-editing biotechnological instrument, with several applications in agriculture. The recent advent of the CRISPR (clustered regularly interspaced short palindromic repeat)/Cas technology can provide important advances in editing the genomes of plant crops including rice, wheat, tomato, maize, and others. In this context, biotechnological advances are essential to overcome such challenges. Modern agriculture is facing several challenges, as food demand increases and climate and dietary patterns change. This Review can aid discussions on the relevance of evaluating off-target modifications for risk assessment of CRISPR-Cas-edited plants. CRISPR-Cas-edited plants showed lower off-target mutation frequencies than conventionally bred plants. Off-targets edits were predominantly detected via biased analysis of predicted off-target sites instead of unbiased genome-wide analysis. Off-target sites had few mismatches (1–3 nt) with the target sequence and were mainly located in protein-coding regions, often in target gene homologues. One study detected the insertion of vector-derived DNA sequences, which is important considering the risk assessment of gene-edited plants. Most observed off-target changes were small insertions or deletions (1–22 bp) or nucleotide substitutions, and large deletions (>100 bp) were rare. Through an extensive literature review we gathered information on CRISPR-Cas off-target edits in plants. Nevertheless, unintended off-target edits can arise that might confer risks when present in gene-edited food crops. Finally, we discuss the future applications of CRISPR technologies in developing herbicide-tolerant crops.ĬRISPR-Cas-based genome editing allows for precise and targeted genetic modification of plants. In this review, we briefly summarize the latest development and breakthroughs of CRISPR technologies in creating herbicide-tolerant crops. Among these, CRISPR technologies, including nuclease systems, base editors, and prime editors, are really promising in creating novel crop germplasms with herbicide tolerance as they are simple, easy to use, and highly efficient. In recent years, the emerging genome technologies, including ZFNs (zinc-finger nucleases), TALENs (transcription activator-like effector nucleases), and CRISPR (clustered regularly interspaced short palindromic repeat), provide us a new way for crop improvement through precise manipulation of endogenous genes in the plant genomes. Development of herbicide-tolerant crops was initially based on conventional plant breeding and transgenic technology. Thus, there is a necessity to develop crop varieties with herbicide resistance or tolerance, which would provide cost-effective tools for helping farmers control weeds in the field. Weeds are one of the biggest problems that modern agriculture is facing worldwide due to the impact they have on crop productivity.

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