Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Zhou, Junming; Luan, Xinchao; Liu, Yixuan; Wang, Lixue; Wang, Jiaxin; Yang, Songnan; Liu, Shuying; Zhang, Jun; Liu, Huijing; Yao, Dan

doi:10.3390/plants12071478

Cited by 12 publications

(3 citation statements)

References 207 publications

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“…As a genetic engineering platform, CRISPR-Cas9 came into use in 2012, and for plants it came into use in 2013. This approach proved to be easy to develop, cost-effective, and versatile in generating targeted gene mutations and alterations for a variety of plant species, from model plants to some economically important crops [35]. The implementation of this approach in editing chloroplast genomes has encountered some problems.…”

Section: Genome Editing Systems Applicable To Chloroplastsmentioning

confidence: 99%

The Search of a Molecular “Swiss Knife” for Chloroplast Genomic Editing

Dorogova,

Sidorchuk

2023

Horticulturae

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In recent years, genome editing methods have become an integral part of the genetic engineering toolset that allows for making targeted changes to plant genomes, both in the case of single-gene mutations and multiplex modifications. These technologies were mostly proven effective for editing nuclear genomes. However, plastids, the best-known example of which is chloroplasts, have their own genome (plastome), which is also available for various genetic manipulations, including editing. Despite the fact that the modification of plastomes represents a very promising task for modern biotechnology, the structure of plastids and the peculiarities of their genome organization require the specific adaptation of genome editing methods. This applies to both the design of genetic constructs and methods of their delivery to plastids. The article provides an overview of the current state of research in the field of plastid genome editing with chloroplasts taken as an example. We consider the possibilities of using programmable genome-editing technologies, analyze their effectiveness, limitations, and problems caused by the structural features of these organelles, and their genome organization. We discuss the results of the first successful experiments in this field and try to assess the prospects for the development of tools and methods for increasing the efficiency and the specificity of this biotechnological platform.

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Section: Genome Editing Systems Applicable To Chloroplastsmentioning

confidence: 99%

The Search of a Molecular “Swiss Knife” for Chloroplast Genomic Editing

Dorogova,

Sidorchuk

2023

Horticulturae

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“…Compared with Cas9, the CRISPR family member Cas12a is more practical and effective. Hence, CRISPR/Cas12a can effectively edit multiple genes because of the specific way that CRISPR RNA (crRNA) functions (Bandyopadhyay et al, 2020;Paul and Montoya, 2020;Zhou et al, 2023b). In 2017, Jiang et al used CRISPR/Cas12a to achieve editing in the soybean FAD2 gene for the first time (Jiang et al, 2017).…”

Section: Application Of Other Crispr Gene Editing Technology In Soybeanmentioning

confidence: 99%

Advances in CRISPR/Cas9-based research related to soybean [Glycine max (Linn.) Merr] molecular breeding

Yao,

Zhou,

Zhang

et al. 2023

Front. Plant Sci.

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Soybean [Glycine max (Linn.) Merr] is a source of plant-based proteins and an essential oilseed crop and industrial raw material. The increase in the demand for soybeans due to societal changes has coincided with the increase in the breeding of soybean varieties with enhanced traits. Earlier gene editing technologies involved zinc finger nucleases and transcription activator-like effector nucleases, but the third-generation gene editing technology uses clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9). The rapid development of CRISPR/Cas9 technology has made it one of the most effective, straightforward, affordable, and user-friendly technologies for targeted gene editing. This review summarizes the application of CRISPR/Cas9 technology in soybean molecular breeding. More specifically, it provides an overview of the genes that have been targeted, the type of editing that occurs, the mechanism of action, and the efficiency of gene editing. Furthermore, suggestions for enhancing and accelerating the molecular breeding of novel soybean varieties with ideal traits (e.g., high yield, high quality, and durable disease resistance) are included.

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“…[ 127 ] In our experience of creating a rapeseed mutant collection by a pooled CRISPR library, the editing efficiency of CRISPR‐Cas9 and the regeneration efficiency of transgenics by tissue culture are two vital limited factors. [ 92 , 128 ] Optimizing the CRISPR‐Cas9 system to improve its editing efficiency in plants, [ 129 ] such as machine learning‐facilitated editing outcome prediction and rational sgRNA design, [ 127 ] will be required in future studies. In addition, novel plasmid delivery methods bypassing tissue culture, such as virus‐based [ 130 , 131 ] and carbon nanocarrier‐based delivery [ 132 , 133 , 134 ] and de novo induction of meristems, [ 135 ] might be used in arrayed CRISPR library screening in the future.…”

Section: Future Perspectivementioning

confidence: 99%

Plant Functional Genomics Based on High‐Throughput CRISPR Library Knockout Screening: A Perspective

He,

Zeng,

2023

Advanced Genetics

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Plant biology studies in the post‐genome era have been focused on annotating genome sequences’ functions. The established plant mutant collections have greatly accelerated functional genomics research in the past few decades. However, most plant genome sequences' roles and the underlying regulatory networks remain substantially unknown. Clustered, regularly interspaced short palindromic repeat (CRISPR)‐associated systems are robust, versatile tools for manipulating plant genomes with various targeted DNA perturbations, providing an excellent opportunity for high‐throughput interrogation of DNA elements’ roles. This study compares methods frequently used for plant functional genomics and then discusses different DNA multi‐targeted strategies to overcome gene redundancy using the CRISPR‐Cas9 system. Next, this work summarizes recent reports using CRISPR libraries for high‐throughput gene knockout and function discoveries in plants. Finally, this work envisions the future perspective of optimizing and leveraging CRISPR library screening in plant genomes' other uncharacterized DNA sequences.

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Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Cited by 12 publications

References 207 publications

The Search of a Molecular “Swiss Knife” for Chloroplast Genomic Editing

The Search of a Molecular “Swiss Knife” for Chloroplast Genomic Editing

Advances in CRISPR/Cas9-based research related to soybean [Glycine max (Linn.) Merr] molecular breeding

Plant Functional Genomics Based on High‐Throughput CRISPR Library Knockout Screening: A Perspective

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