Design of Guide RNA for CRISPR/Cas Plant Genome Editing

Геращенков, Г. А.; Рожнова, Н. А.; Kuluev, B. R.; Kiryanova, O.Yu.; Gumerova, G. R.; Knyazev, A. V.; Вершинина, З. Р.; Михайлова, Е. В.; Чемерис, Д.А.; Matniyazov, R. T.; Gubaydullin, I. M.; Баймиев, Ал. Х.; Чемерис, А. В.

doi:10.1134/s0026893320010069

Cited by 33 publications

(18 citation statements)

References 83 publications

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“…Generally, these tools exploit a similar algorithm to design gRNA [ 255 ]. However, some characteristics related to the gRNA spacer region (e.g., number and type of mismatch, GC-content, length, and indels) and the selection of specific Cas-nucleases vary between these tools [ 256 ].…”

Section: Genetic Engineering Approaches In Cannabismentioning

confidence: 99%

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Hesami

Baiton

Alizadeh

et al. 2021

IJMS

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For a long time, Cannabis sativa has been used for therapeutic and industrial purposes. Due to its increasing demand in medicine, recreation, and industry, there is a dire need to apply new biotechnological tools to introduce new genotypes with desirable traits and enhanced secondary metabolite production. Micropropagation, conservation, cell suspension culture, hairy root culture, polyploidy manipulation, and Agrobacterium-mediated gene transformation have been studied and used in cannabis. However, some obstacles such as the low rate of transgenic plant regeneration and low efficiency of secondary metabolite production in hairy root culture and cell suspension culture have restricted the application of these approaches in cannabis. In the current review, in vitro culture and genetic engineering methods in cannabis along with other promising techniques such as morphogenic genes, new computational approaches, clustered regularly interspaced short palindromic repeats (CRISPR), CRISPR/Cas9-equipped Agrobacterium-mediated genome editing, and hairy root culture, that can help improve gene transformation and plant regeneration, as well as enhance secondary metabolite production, have been highlighted and discussed.

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Section: Genetic Engineering Approaches In Cannabismentioning

confidence: 99%

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Hesami

Baiton

Alizadeh

et al. 2021

IJMS

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“…Another major drawback of the CRISPR-Cas system is its non-specificity. In this case, Cas cleaves DNA at non-target sites that are not complementary of single guide RNA [ 172 ]. This drawback impedes CRISPR-Cas potential applications, particularly when genome alteration needs to be precise, as in the case of gene therapy.…”

Section: Constraints and Solutions Related To Crispr-cas Genome Editingmentioning

confidence: 99%

“…In plants, the specificity of the CRISPR-Cas system has been estimated by deliberate non-target investigation [ 174 ]. For RNPs, non-target alterations were hardly recognized by thorough sequencing, indicating that RNPs enhance the specificity of the editing system [ 172 ]. However, no study has been reported on Cas specificity in plants.…”

Section: Constraints and Solutions Related To Crispr-cas Genome Editingmentioning

confidence: 99%

From Traditional Breeding to Genome Editing for Boosting Productivity of the Ancient Grain Tef [Eragrostis tef (Zucc.) Trotter]

Numan

Khan

Asaf

et al. 2021

Plants

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Tef (Eragrostis tef (Zucc.) Trotter) is a staple food crop for 70% of the Ethiopian population and is currently cultivated in several countries for grain and forage production. It is one of the most nutritious grains, and is also more resilient to marginal soil and climate conditions than major cereals such as maize, wheat and rice. However, tef is an extremely low-yielding crop, mainly due to lodging, which is when stalks fall on the ground irreversibly, and prolonged drought during the growing season. Climate change is triggering several biotic and abiotic stresses which are expected to cause severe food shortages in the foreseeable future. This has necessitated an alternative and robust approach in order to improve resilience to diverse types of stresses and increase crop yields. Traditional breeding has been extensively implemented to develop crop varieties with traits of interest, although the technique has several limitations. Currently, genome editing technologies are receiving increased interest among plant biologists as a means of improving key agronomic traits. In this review, the potential application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (CRISPR-Cas) technology in improving stress resilience in tef is discussed. Several putative abiotic stress-resilient genes of the related monocot plant species have been discussed and proposed as target genes for editing in tef through the CRISPR-Cas system. This is expected to improve stress resilience and boost productivity, thereby ensuring food and nutrition security in the region where it is needed the most.

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“…This is because the specificity of sgRNAs are highly dependent on the so-called "seed region" of crRNAs, which in the case of Cas9, encompasses approximately 8-12 nucleotides upstream of the PAM. Mismatches of 2 or more nucleotides in this sequence of the protospacer generally prevent Cas9 cleavage (Tang et al, 2018;Young et al, 2019;Gerashchenkov et al, 2020). This means that possible off-target mutations are highly predictable and can very easily be minimized through the careful selection of target sites.…”

Section: Potential For Off-target Effects In Plantsmentioning

confidence: 99%

Genetic Variation and Unintended Risk in the Context of Old and New Breeding Techniques

Singer

Laurie

Bilichak

et al. 2021

Critical Reviews in Plant Sciences

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For thousands of years, humans have been improving crops to better suit their needs. These enhancements are driven by changes in the genetic makeup of the plant. While this was initially unintentional, there has been a steady push to increase the pace and precision of crop breeding, something that has occurred alongside a growing understanding of genetics and an escalating capacity to thoroughly assess genomes at the molecular level. With the advent and rapid uptake of molecular breeding techniques, such as transgenics and genome editing over the past few decades, there has been much trepidation regarding the possibility of off-target effects derived from unanticipated mutations at loci other than those intended for alteration, and the unintended risks that this might confer. These concerns persist regardless of the fact that a growing number of studies indicate that the occurrence of off-target mutations derived from newer biotechnological breeding techniques are negligible compared to what is observed with many conventional breeding approaches, and even spontaneously from one generation to the next. Given the impending food security crisis that we are facing in the short-term, there is a critical need to implement a wide range of breeding tools as a means of meeting growing demand, withstanding climate change-related pressures, increasing nutrition, and providing environmental benefits. While food safety is clearly of the utmost importance, now is certainly not the time to prevent the use of particular breeding technologies based on unfounded doubts. Therefore, in this review, we attempt to shed light on these apprehensions by putting purported "risks" into the context of plant breeding as a whole by comparing frequencies of spontaneous mutations with those (both anticipated and unanticipated) that occur through various conventional and biotechnological breeding approaches, including transgenics and genome editing. We then consider how these changes may, or may not, translate into unanticipated risk, and discuss the current global regulatory asynchrony surrounding genome edited crops.

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Design of Guide RNA for CRISPR/Cas Plant Genome Editing

Cited by 33 publications

References 83 publications

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

From Traditional Breeding to Genome Editing for Boosting Productivity of the Ancient Grain Tef [Eragrostis tef (Zucc.) Trotter]

Genetic Variation and Unintended Risk in the Context of Old and New Breeding Techniques

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