Cas14a1-mediated nucleic acid detectifon platform for pathogens

Ge, Xiaolin; Meng, Tian; Tan, Xiao; Wei, Yangdao; Tao, Zhenzhen; Yang, Zhiqing; Song, Fengge; Wang, Peng; Wan, Yi

doi:10.1016/j.bios.2021.113350

Cited by 51 publications

(24 citation statements)

References 33 publications

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“…Cas14, which has only 400 to 700 amino acids, may target ssDNA and induce cleavage without requiring a specific sequence ( Harrington et al, 2018 ). Up to now, the applications of Cas9, Cpf1 variants, Cas13a, and Cas14 are mainly focused on nucleic acid testing ( Gootenberg et al, 2017 ; Ge et al, 2021 ) or mammalian ( Gao et al, 2017 ; Walton et al, 2020 ) and plant cells ( Abudayyeh et al, 2017 ). Based on their advantages, they will have promising potentials to be developed in non-conventional yeasts.…”

Section: Discussionmentioning

confidence: 99%

Advances and Opportunities of CRISPR/Cas Technology in Bioengineering Non-conventional Yeasts

Shan

Dai

Wang

2021

Front. Bioeng. Biotechnol.

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Non-conventional yeasts have attracted a growing interest on account of their excellent characteristics. In recent years, the emerging of CRISPR/Cas technology has improved the efficiency and accuracy of genome editing. Utilizing the advantages of CRISPR/Cas in bioengineering of non-conventional yeasts, quite a few advancements have been made. Due to the diversity in their genetic background, the ways for building a functional CRISPR/Cas system of various species non-conventional yeasts were also species-specific. Herein, we have summarized the different strategies for optimizing CRISPR/Cas systems in different non-conventional yeasts and their biotechnological applications in the construction of cell factories. In addition, we have proposed some potential directions for broadening and improving the application of CRISPR/Cas technology in non-conventional yeasts.

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Section: Discussionmentioning

confidence: 99%

Advances and Opportunities of CRISPR/Cas Technology in Bioengineering Non-conventional Yeasts

Shan

Dai

Wang

2021

Front. Bioeng. Biotechnol.

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“…[8] Unlike Cas13abased RNAd etection, [16] Cas14a1 had ah igh discrimination capacity against single mismatches in the target RNA. [17] CRISPR-Cas II effectors with trans-cleavage activity (more detail functional information of CRISPR-Cas II effectors shown Table S5), including Cas12, [18] Cas13, [19] and Cas14, [20] have been used as biosensing platforms. [21] Numerous works have combined Cas effectors-based methods with isothermal amplification to obtain high detection sensitivity, [22] such as those obtained by recombinase polymerase amplification (RPA) [23] and strand displacement amplification.…”

Section: Methodsmentioning

confidence: 99%

trans Single‐Stranded DNA Cleavage via CRISPR/Cas14a1 Activated by Target RNA without Destruction

et al. 2021

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As aC RISPR-Cas system (clustered regularly interspaced short palindromic repeats and CRISPR associated proteins), Cas14a1 can cis/trans cleave single-stranded DNA (ssDNA). Here,w ed escribe an unreported capacity of Cas14a1:R NA can trigger the trans ssDNAc leavage.T his Cas14a1-based RNA-activated detection platform (Amplification, Transcription, Cas14a1-based RNA-activated trans ssDNAc leavage,A TCas-RNA) has an outstanding specificity for the detection of target RNAs with point mutation resolution, which is better than that of the Cas14a1-based ssDNAactivation. Using ATCas-RNAv ia af luorophore quencherlabeled ssDNAr eporter (FQ), we were able to detect 1aM pathogenic nucleic acid within 1h,and achieve 100 %accuracy with 25 milk samples.This platform can serve as anew tool for high-efficiency nucleic acid diagnostics.Importantly,this work can expand our understanding of Cas14a1 and inspire further mechanisms and applications of Class-2 Cas systems.

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“…Ge et al [ 163 ] also used the function of Cas14a to establish a Cas14a1-mediated nucleic acid detection platform (CMP) for the rapid detection of pathogens in milk samples. This promoted the development of pathogen detection in the field of food safety.…”

Section: Application Of Crispr-cas For the Detection Of Bacterial Infectionsmentioning

confidence: 99%

“…The Cas14a1-sgRNA complex binds and cleaves the target ssDNA, activating the collateral cleavage of the nonspecific ssDNA in the presence of fluorescent and quenching agents. In addition, they used CMP and qPCR simultaneously for the quantitative detection of S. pyogenes and S. Typhi in samples, and the results showed that CMP provided higher sensitivity than qPCR [ 163 ].…”

Section: Application Of Crispr-cas For the Detection Of Bacterial Infectionsmentioning

confidence: 99%

Engineered CRISPR-Cas systems for the detection and control of antibiotic-resistant infections

Battalapalli

Hakeem

et al. 2021

J Nanobiotechnol

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Antibiotic resistance is spreading rapidly around the world and seriously impeding efforts to control microbial infections. Although nucleic acid testing is widely deployed for the detection of antibiotic resistant bacteria, the current techniques—mainly based on polymerase chain reaction (PCR)—are time-consuming and laborious. There is an urgent need to develop new strategies to control bacterial infections and the spread of antimicrobial resistance (AMR). The CRISPR-Cas system is an adaptive immune system found in many prokaryotes that presents attractive opportunities to target and edit nucleic acids with high precision and reliability. Engineered CRISPR-Cas systems are reported to effectively kill bacteria or even revert bacterial resistance to antibiotics (resensitizing bacterial cells to antibiotics). Strategies for combating antimicrobial resistance using CRISPR (i.e., Cas9, Cas12, Cas13, and Cas14) can be of great significance in detecting bacteria and their resistance to antibiotics. This review discusses the structures, mechanisms, and detection methods of CRISPR-Cas systems and how these systems can be engineered for the rapid and reliable detection of bacteria using various approaches, with a particular focus on nanoparticles. In addition, we summarize the most recent advances in applying the CRISPR-Cas system for virulence modulation of bacterial infections and combating antimicrobial resistance. Graphical Abstract

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Cas14a1-mediated nucleic acid detectifon platform for pathogens

Cited by 51 publications

References 33 publications

Advances and Opportunities of CRISPR/Cas Technology in Bioengineering Non-conventional Yeasts

Advances and Opportunities of CRISPR/Cas Technology in Bioengineering Non-conventional Yeasts

trans Single‐Stranded DNA Cleavage via CRISPR/Cas14a1 Activated by Target RNA without Destruction

Engineered CRISPR-Cas systems for the detection and control of antibiotic-resistant infections

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