Tools and targets: The dual role of plant viruses in CRISPR–Cas genome editing

Uranga, Mireia; Daròs, José‐Antonio

doi:10.1002/tpg2.20220

Cited by 19 publications

(13 citation statements)

References 228 publications

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“…Plant viral vectors hold great potential for accelerated genome editing and crop improvement due to their modest mechanism of delivery. Plants can be directly inoculated with viral vectors harboring genome editing components, and it can only take between 3–7 days for successful viral genome editing to take place allowing for rapid screening of a large quantity of plants [ 109 ]. The modification of the CenH3 could potentially affect plant fitness and reduce recovery of potential haploid inducer lines from in vitro regeneration.…”

Section: Prospects For Doubled Haploids In Sunflowermentioning

confidence: 99%

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Accelerated Breeding for Helianthus annuus (Sunflower) through Doubled Haploidy: An Insight on Past and Future Prospects in the Era of Genome Editing

Mabuza

Mchunu

Crampton

et al. 2023

Plants

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The aim of any breeding process is to fully express the targeted, superior/desirable parent characteristic in the progeny. Hybrids are often used in this dynamic, and complex process for which homozygous parents—which may require up to eight generations of back crossing and selection—are required. Doubled haploid (DH) technologies can facilitate the production of true breeding lines faster and in a more efficient manner than the traditional back crossing and selection strategies. Sunflower is the third most important oilseed crop in the world and has no available double haploid induction procedure/technique that can be efficiently used in breeding programs. A reproducible and efficient doubled haploid induction method would be a valuable tool in accelerating the breeding of new elite sunflower varieties. Although several attempts have been made, the establishment of a sunflower doubled haploid induction protocol has remained a challenge owing recalcitrance to in vitro culture regeneration. Approaches for haploid development in other crops are often cultivar specific, difficult to reproduce, and rely on available tissue culture protocols—which on their own are also cultivar and/or species specific. As an out-crossing crop, the lack of a double haploid system limits sunflower breeding and associated improvement processes, thereby delaying new hybrid and trait developments. Significant molecular advances targeting genes, such as the centromeric histone 3 (CenH3) and Matrilineal (MTL) gene with CRISPR/Cas9, and the successful use of viral vectors for the delivery of CRISPR/Cas9 components into plant cells eliminating the in vitro culture bottleneck, have the potential to improve double haploid technology in sunflower. In this review, the different strategies, their challenges, and opportunities for achieving doubled haploids in sunflower are explored.

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Section: Prospects For Doubled Haploids In Sunflowermentioning

confidence: 99%

“…The modification of the CenH3 could potentially affect plant fitness and reduce recovery of potential haploid inducer lines from in vitro regeneration. Virus vectors can deliver genome editing reagents to mature plants, and editing can occur before the onset of plant growth-limiting effects [ 109 ].…”

Section: Prospects For Doubled Haploids In Sunflowermentioning

confidence: 99%

Accelerated Breeding for Helianthus annuus (Sunflower) through Doubled Haploidy: An Insight on Past and Future Prospects in the Era of Genome Editing

Mabuza

Mchunu

Crampton

et al. 2023

Plants

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“…CRITICAL: A variety of Cas nuclease effectors have been reported (Uranga and Daròs, 2022). This protocol has been validated using a N. benthamiana transgenic line that constitutively expresses a human codon-optimized version of Streptococcus pyogenes Cas9 fused to a nuclear localization signal (NLS) at the carboxyl terminus.…”

Section: Before You Beginmentioning

confidence: 99%

“…Protocols and computational tools were reported for CRISPR/Cas genome editing [3][4][5] . Viral vectors are a promising alternative for the transient delivery of the CRISPR-Cas system in many biological systems including plants, following the so-called virus-induced genome editing (VIGE) approach [6][7][8] . The systemic spread of the viral infection promotes an accumulation of the editing components within plant tissues.…”

Section: Before You Beginmentioning

confidence: 99%

Heritable CRISPR-Cas9 editing of plant genomes using RNA virus vectors

Uranga

Aragonés

Daròs

et al. 2022

Preprint

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Viral vectors hold enormous potential for genome editing in plants as transient delivery vehicles of CRISPR-Cas components. Here, we describe a protocol to assemble plant viral vectors for single guide RNA (sgRNA) delivery. The obtained viral constructs are based on compact T-DNA binary vectors of the pLX series and are delivered into Cas9-expressing plants through agroinoculation. This approach allows rapidly assessing sgRNA design for plant genome targeting, as well as the recovery of progeny with heritable mutations at targeted loci.

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“…Traditional approaches for the delivery of editing components rely on the transformation protocols ( Agrobacterium and Biolistic) or transient delivery to the protoplasts. However, both techniques are time-consuming, laborious, and also raise some legal concerns [ 9 ]; thus, advanced and reliable techniques are direly needed. One such technique could be the use of plant virus-derived vectors which have already been used for multiple purposes including the production of useful proteins [ 10 ], etc.…”

Section: Introductionmentioning

confidence: 99%

Plant Virus-Derived Vectors for Plant Genome Engineering

Mahmood

Naqvi

Rahman

et al. 2023

Viruses

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Advances in genome engineering (GE) tools based on sequence-specific programmable nucleases have revolutionized precise genome editing in plants. However, only the traditional approaches are used to deliver these GE reagents, which mostly rely on Agrobacterium-mediated transformation or particle bombardment. These techniques have been successfully used for the past decades for the genetic engineering of plants with some limitations relating to lengthy time-taking protocols and transgenes integration-related regulatory concerns. Nevertheless, in the era of climate change, we require certain faster protocols for developing climate-smart resilient crops through GE to deal with global food security. Therefore, some alternative approaches are needed to robustly deliver the GE reagents. In this case, the plant viral vectors could be an excellent option for the delivery of GE reagents because they are efficient, effective, and precise. Additionally, these are autonomously replicating and considered as natural specialists for transient delivery. In the present review, we have discussed the potential use of these plant viral vectors for the efficient delivery of GE reagents. We have further described the different plant viral vectors, such as DNA and RNA viruses, which have been used as efficient gene targeting systems in model plants, and in other important crops including potato, tomato, wheat, and rice. The achievements gained so far in the use of viral vectors as a carrier for GE reagent delivery are depicted along with the benefits and limitations of each viral vector. Moreover, recent advances have been explored in employing viral vectors for GE and adapting this technology for future research.

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Tools and targets: The dual role of plant viruses in CRISPR–Cas genome editing

Cited by 19 publications

References 228 publications

Accelerated Breeding for Helianthus annuus (Sunflower) through Doubled Haploidy: An Insight on Past and Future Prospects in the Era of Genome Editing

Accelerated Breeding for Helianthus annuus (Sunflower) through Doubled Haploidy: An Insight on Past and Future Prospects in the Era of Genome Editing

Heritable CRISPR-Cas9 editing of plant genomes using RNA virus vectors

Plant Virus-Derived Vectors for Plant Genome Engineering

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