Genome editing in plants with MAD7 nuclease

Lin, Qiupeng; Zhu, Zixu; Liu, Guanwen; Sun, Chao; Lin, Dexing; Xue, Chenxiao; Li, Shengnan; Zhang, Dandan; Gao, Caixia; Wang, Yanpeng; Qiu, Jian

doi:10.1016/j.jgg.2021.04.003

Cited by 29 publications

(40 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Because MAD7 "is royalty-free for both academic and commercial research and development use" (Inscripta, 2021;Price et al, 2020;Rojek et al, 2022) it serves as ideal tool for strain engineering in an industrial and commercial environment. MAD7-mediated genome engineering has been demonstrated both in eukaryotic systems including fungi (Jarczynska et al, 2021), plants (Lin et al, 2021), vertebrate and mammalian systems (Liu et al, 2020;Wierson et al, 2019), as well as in the prokaryotic organisms E. coli and Bacillus subtilis (Price et al, 2020). Previous use of MAD7 in E. coli focused on screening or nuclease engineering (Dewachter et al, 2022;Liu et al, 2019).…”

Section: Resultsmentioning

confidence: 99%

A MAD7-based genome editing system for E. coli

Mund

Weber

Degreif

et al. 2022

Preprint

View full text Add to dashboard Cite

A broad variety of biomolecules is industrially produced in bacteria and yeasts. These microbial expression hosts can be optimized through genomic engineering using CRISPR tools. Here, we designed and characterized such a modular genome editing system based on the Cas12a-like RNA guided nuclease MAD7 in E. coli. Our system enables the efficient generation of SNPs or gene deletions and can directly be used with donor DNA from benchtop DNA assembly to increase throughput. We combined multiple edits to engineer an E. coli strain with reduced overflow metabolism and increased plasmid yield, highlighting the versatility and industrial applicability of our approach.

show abstract

Section: Resultsmentioning

confidence: 99%

A MAD7-based genome editing system for E. coli

Mund

Weber

Degreif

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Engineered Cas MAD7-RR, MAD7-RVR, and M-AFID (MAD7-APOBEC fusion-induced deletion) increase the targeting range of MAD7 by creating predictable deletions from 5′-deaminated Cs to the MAD7-cleavage site. This new CRISPR-MAD7 system has an efficiency of up to 65.6%, as demonstrated in mutant rice and wheat plants ( Lin et al, 2021 ). MAD7 can expand the CRISPR toolbox for genome engineering due to its highly efficient target to gene disruption and insertions, different protospacer adjacent motifs, and small-guide RNA requirements ( Liu et al, 2020 ).…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 92%

Clustered Regularly Interspaced Short Palindromic Repeats-Associated Protein System for Resistance Against Plant Viruses: Applications and Perspectives

Silva

Fontes

2022

Front. Plant Sci.

View full text Add to dashboard Cite

Different genome editing approaches have been used to engineer resistance against plant viruses. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas; CRISPR/Cas) systems to create pinpoint genetic mutations have emerged as a powerful tool for molecular engineering of plant immunity and increasing resistance against plant viruses. This review presents (i) recent advances in engineering resistance against plant viruses by CRISPR/Cas and (ii) an overview of the potential host factors as targets for the CRISPR/Cas system-mediated broad-range resistance and immunity. Applications, challenges, and perspectives in enabling the CRISPR/Cas system for crop protection are also outlined.

show abstract

“…If no knock-in donor template is present, Cas12a target sites can be cut and repaired by NHEJ several times until a large deletion occurs and deletes the PAM, preventing further cleavage. Consequently, 3-30-nt deletions are frequently observed with Cas12a [10,13,[30][31][32][33], whereas Cas9 mostly induces 1-2-nt indels [13]. Although this property may favor Cas12a for gene knock-outs [19,33], preventing unwanted large deletions may be required for clinical use of Cas12a.…”

Section: Comparison Of Crispr-cas12a and Crispr-cas9mentioning

confidence: 99%

Current and Prospective Applications of CRISPR-Cas12a in Pluricellular Organisms

Khan

Sallard

2022

Mol Biotechnol

View full text Add to dashboard Cite

CRISPR-Cas systems play a critical role in the prokaryotic adaptive immunity against mobile genetic elements, such as phages and foreign plasmids. In the last decade, Cas9 has been established as a powerful and versatile gene editing tool. In its wake, the novel RNA-guided endonuclease system CRISPR-Cas12a is transforming biological research due to its unique properties, such as its high specificity or its ability to target T-rich motifs, to induce staggered double-strand breaks and to process RNA arrays. Meanwhile, there is an increasing need for efficient and safe gene activation, repression or editing in pluricellular organisms for crop improvement, gene therapy, research model development, and other goals. In this article, we review CRISPR-Cas12a applications in pluricellular organisms and discuss how the challenges characteristic of these complex models, such as vectorization or temperature variations in ectothermic species, can be overcome.

show abstract

Genome editing in plants with MAD7 nuclease

Cited by 29 publications

References 53 publications

A MAD7-based genome editing system for E. coli

A MAD7-based genome editing system for E. coli

Clustered Regularly Interspaced Short Palindromic Repeats-Associated Protein System for Resistance Against Plant Viruses: Applications and Perspectives

Current and Prospective Applications of CRISPR-Cas12a in Pluricellular Organisms

Contact Info

Product

Resources

About