The CRISPR-Cas toolbox and gene editing technologies

Liu, Guanwen; Lin, Qiupeng; Jin, Shuai; Gao, Caixia

doi:10.1016/j.molcel.2021.12.002

Cited by 204 publications

(147 citation statements)

References 172 publications

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“…However, their mutation ratios are low, and the mutation sites are random and unknown. The CRISPR/Cas9 system can easily produce target gene mutation, and has been widely used in plant mutant creation, especially for reverse genetics in revealing target gene function and regulation [6][7][8][9][10]. The Cas9 protein functions as a nuclease and is guided to the target site by the engineered single guide RNA (gRNA) including 20 specific nucleotides of the selected gene to determine the site-specific targeting.…”

Section: Discussionmentioning

confidence: 99%

“…The reverse genetics can reveal the gene function through the mutation or expression regulation of target gene [1,2]. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated (Cas) system can easily mutate and edit the target gene, and has been widely used in plant sciences and crop improvement [6][7][8][9][10]. The mutants induced by CRISPR/Cas system usually have many heterozygous and bi-allelic mutant lines, which can produce the homozygous mutant plants in their offspring segregation population [11][12][13][14][15][16][17].…”

Section: Introductionmentioning

confidence: 99%

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Application of Allele Specific PCR in Identifying Offspring Genotypes of Bi-Allelic SbeIIb Mutant Lines in Rice

Jiang

Ren

Wang

et al. 2022

Plants

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Bi-allelic mutant lines induced by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated (Cas) systems are important genetic materials. It is very important to establish a rapid and cheap method in identifying homozygous mutant plants from offspring segregation populations of bi-allelic mutant lines. In this study, the offspring genotypes of rice bi-allelic starch branching enzyme IIb mutant lines were identified using the allele specific PCR (AS-PCR) method. The target sequences of two alleles were aligned from their 5′ to 3′ ends, and the first different bases were used as the 3′ ends of mismatch primers. Another mismatched base was introduced at the third nucleotide from the 3′ end of mismatch primer. The PCR reaction mixture and amplification program were optimized according to the differences of mutation target sequence and mismatch primers. The offspring plant genotypes of bi-allelic mutant lines could be accurately identified using the amplified DNA fragments by agarose gel electrophoresis. This study could provide a method reference for the rapid screening of homozygous mutant plants from offspring segregation population of heterozygous and bi-allelic mutant lines.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Application of Allele Specific PCR in Identifying Offspring Genotypes of Bi-Allelic SbeIIb Mutant Lines in Rice

Jiang

Ren

Wang

et al. 2022

Plants

View full text Add to dashboard Cite

show abstract

“…These can be achieved through gene editing or silencing using DNA or RNA-based tools. DNA-based tools, which usually target gene transcription, include the use of Zinc Finger Nucleases (ZFN), Transcription Activator-Like Effector Nucleases (TALEN) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) methods [ 34 ], while RNA-based tools, such as RNA interference and anti-sense oligonucleotides, and even more recent RNA editing methods [ 35 ], exist for targeting gene translation, although the effects are usually transient. Observing the natural history of IRDs and conducting long-term investigations of therapeutics in these models may be challenging.…”

Section: Genetic Methods For Ird Nhp Model Creationmentioning

confidence: 99%

Developing Non-Human Primate Models of Inherited Retinal Diseases

Seah

Goh

Chan

et al. 2022

Genes

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Inherited retinal diseases (IRDs) represent a genetically and clinically heterogenous group of diseases that can eventually lead to blindness. Advances in sequencing technologies have resulted in better molecular characterization and genotype–phenotype correlation of IRDs. This has fueled research into therapeutic development over the recent years. Animal models are required for pre-clinical efficacy assessment. Non-human primates (NHP) are ideal due to the anatomical and genetic similarities shared with humans. However, developing NHP disease to recapitulate the disease phenotype for specific IRDs may be challenging from both technical and cost perspectives. This review discusses the currently available NHP IRD models and the methods used for development, with a particular focus on gene-editing technologies.

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“…Since the Cas proteins showed remarkable diversity, CRISPR systems were divided into two classes according to the effector modules composed of multiple Cas proteins (class 1, involving types I, III, and IV) or single multidomain protein (class 2, involving types II, V, and VI) ( Makarova et al, 2020 ). Although class 1 systems extensively exist in microorganisms, their applications were limited by the need for multi-subunit effectors ( Liu et al, 2021a ). Thanks to the efforts on the discovery of novel systems, highly efficient and simplified CRISPR-based biosensing methods were designed.…”

Section: Crispr-based Biosensing Techniquesmentioning

confidence: 99%

Powerful CRISPR-Based Biosensing Techniques and Their Integration With Microfluidic Platforms

Chen

et al. 2022

Front. Bioeng. Biotechnol.

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In the fight against the worldwide pandemic coronavirus disease 2019 (COVID-19), simple, rapid, and sensitive tools for nucleic acid detection are in urgent need. PCR has been a classic method for nucleic acid detection with high sensitivity and specificity. However, this method still has essential limitations due to the dependence on thermal cycling, which requires costly equipment, professional technicians, and long turnover times. Currently, clustered regularly interspaced short palindromic repeats (CRISPR)-based biosensors have been developed as powerful tools for nucleic acid detection. Moreover, the CRISPR method can be performed at physiological temperature, meaning that it is easy to assemble into point-of-care devices. Microfluidic chips hold promises to integrate sample processing and analysis on a chip, reducing the consumption of sample and reagent and increasing the detection throughput. This review provides an overview of recent advances in the development of CRISPR-based biosensing techniques and their perfect combination with microfluidic platforms. New opportunities and challenges for the improvement of specificity and efficiency signal amplification are outlined. Furthermore, their various applications in healthcare, animal husbandry, agriculture, and forestry are discussed.

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The CRISPR-Cas toolbox and gene editing technologies

Cited by 204 publications

References 172 publications

Application of Allele Specific PCR in Identifying Offspring Genotypes of Bi-Allelic SbeIIb Mutant Lines in Rice

Application of Allele Specific PCR in Identifying Offspring Genotypes of Bi-Allelic SbeIIb Mutant Lines in Rice

Developing Non-Human Primate Models of Inherited Retinal Diseases

Powerful CRISPR-Based Biosensing Techniques and Their Integration With Microfluidic Platforms

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