The Rise of the CRISPR/Cpf1 System for Efficient Genome Editing in Plants

Alok, Anshu; Sandhya, Dulam; Jogam, Phanikanth; Rodrigues, Vandasue; Bhati, Kaushal Kumar; Sharma, Himanshu; Kumar, Jitendra

doi:10.3389/fpls.2020.00264

Cited by 82 publications

(51 citation statements)

References 58 publications

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“…Genome editing using Cpf1 requires higher temperatures for efficient editing efficiency. The Lachnospiraceae bacterium Cpf1 showed less editing efficiency at 22 °C, whereas it showed 100% editing efficiency at 28 °C [ 88 ]. CRISPR/Cas9-mediated genome editing of citrus also showed higher editing efficiency when plants were subjected to heat stress at 37 °C [ 87 ].…”

Section: Main Textmentioning

confidence: 99%

The present and potential future methods for delivering CRISPR/Cas9 components in plants

Sandhya

Jogam

Allini

et al. 2020

Journal of Genetic Engineering and Biotechnology

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Background CRISPR/Cas9 genome editing technology is a DNA manipulation tool for trait improvement. This technology has been demonstrated and successfully applied to edit the genome in various species of plants. The delivery of CRISPR/Cas9 components within rigid plant cells is very crucial for high editing efficiency. Here, we insight the strengths and weaknesses of each method of delivery. Main text The mutation efficiency of genome editing may vary and affected by different factors. Out of various factors, the delivery of CRISPR/Cas9 components into cells and genome is vital. The way of delivery defines whether the edited plant is transgenic or transgene-free. In many countries, the transgenic approach of improvement is a significant limitation in the regulatory approval of genetically modified crops. Gene editing provides an opportunity for generating transgene-free edited genome of the plant. Nevertheless, the mode of delivery of the CRISPR/Cas9 component is of crucial importance for genome modification in plants. Different delivery methods such as Agrobacterium -mediated, bombardment or biolistic method, floral-dip, and PEG-mediated protoplast are frequently applied to crops for efficient genome editing. Conclusion We have reviewed different delivery methods with prons and cons for genome editing in plants. A novel nanoparticle and pollen magnetofection-mediated delivery systems which would be very useful in the near future. Further, the factors affecting editing efficiency, such as the promoter, transformation method, and selection pressure, are discussed in the present review.

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Section: Main Textmentioning

confidence: 99%

The present and potential future methods for delivering CRISPR/Cas9 components in plants

Sandhya

Jogam

Allini

et al. 2020

Journal of Genetic Engineering and Biotechnology

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“…Step: expression of CrRNAs from the transcription of CRISPR array which also involves the expression of the Cas9 protein; fashion to generate blunt ends 84 . CRISPR/Cpf1 has been used in many plants 85 . Furthermore, it is necessary to insert or delete the nucleotide sequences for the improvement of crop traits.…”

Section: Developmental History Of Talens Is Incomplete Without Intermentioning

confidence: 99%

Conventional Breeding, Molecular Breeding and Speed Breeding; Brave Approaches to Revamp the Production of Cereal Crops

Haroon¹,

Mm²,

Farooq³

et al. 2020

Preprint

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Conventional plant breeding methods exploit already existing genomic variation in plants to develop a variety in 8 to 10 years, which can decrease the genetic variability of the plant’s genome. The ever-increasing food demand for cereals crops cannot be met by traditional breeding methods. In order to increase food production in less time, there is a dire need to improve breeding methods. Several conventional and molecular breeding methods are being used to improve the crops traits. Molecular researchers have developed new genome editing tools like CRISPR/Cas9, CRISPR/Cpf1, prime editing, base editing, dcas9 epigenetic modification, and several other transgene-free genomes editing approaches. These genome editing tools can improve the desired traits precisely and efficiently. Moreover, a newly developed breeding method “Speed Breeding” has revolutionized the agriculture by shortening the crop cycle. It can produce 5-6 generations of cereals in a year. In this review, we have summarized all these conventional and molecular breeding approaches to improve cereal crops.

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“…Compared to CRISPR/Cas9 system, Cpf1 endonuclease has a smaller molecular weight, and requires shorter CRISPR RNA (crRNA) 37,38 . These advantages of CRISPR/Cpf1 system could help to reduce the overall size of the plant transformation vector, making it more suitable for multiplexed genome editing for plants 39 . However, the CRISPR/Cpf1 system has not been applied in Orchidaceae so far.…”

Section: Assessment Of Dpⅱ-cpf1 Multiplex Genome Editing System In Phmentioning

confidence: 99%

Efficient Multiplex Genome Editing Tools identified by Protoplast Technology inPhalaenopsis

Xia

Zhang

Liu

et al. 2020

Preprint

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Phalaenopsis orchids are popular ornamental plants worldwide. The application of the efficient multiplex genome editing tools in Phalaenopsis , will greatly accelerate the development of orchid gene function and breeding research. In this study, we establish a fast and convenient Phalaenopsis protoplast platform for the identification of functional genome editing tools. Two multiplex genome editing tools, PTG-Cas9 (PTG, polycistronic tRNA gRNA) system and PTGm-Cas9 (PTG-Cas9 system with modified sgRNA structure) system are designed to edit PDS gene of commercial Phalaenopsis ST166 at four target sites. We find that both PTG-Cas9 and PTGm-Cas9 system are functional in Phalaenopsis , and the PTGm-Cas9 system with modified sgRNA has a higher editing efficiency than PTG-Cas9 system. Further, we design another multiplex genome editing tool, termed as DPⅡ-Cpf1 system (dual Pol II promoter to drive the expression of Cpf1 endonuclease and crRNA), to edit PDS gene of Phalaenopsis at four target sites likewise. All the four targets are efficiently edited by DPⅡ-Cpf1 system, and the total mutation rate is about 3 times higher than that of PTGm-Cas9 system. Taken together, using the Phalaenopsis protoplast platform, we successfully establish two efficient multiplex genome editing tools for Phalaenopsis research, PTGm-Cas9 and DPⅡ-Cpf1. The multiplex genome editing tools established in this study have great application potentials in efficiently constructing large-scale knockout mutant libraries of orchid and speeding up orchid precise breeding.

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The Rise of the CRISPR/Cpf1 System for Efficient Genome Editing in Plants

Cited by 82 publications

References 58 publications

The present and potential future methods for delivering CRISPR/Cas9 components in plants

The present and potential future methods for delivering CRISPR/Cas9 components in plants

Conventional Breeding, Molecular Breeding and Speed Breeding; Brave Approaches to Revamp the Production of Cereal Crops

Efficient Multiplex Genome Editing Tools identified by Protoplast Technology inPhalaenopsis

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