An integrated CRISPR Bombyx mori genome editing system with improved efficiency and expanded target sites

Ma, Sanyuan; Liu, Yue; Liu, Yuanyuan; Chang, Jiasong; Zhang, Tong; Wang, Xiaogang; Shi, Run; Lu, Wei; Xia, Xiaojuan; Zhao, Ping; Xia, Qingyou

doi:10.1016/j.ibmb.2017.02.003

Cited by 34 publications

(36 citation statements)

References 35 publications

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“…Ma et al . (2017b) introduced SaCas9 and AsCpf1, in which PAM sites are NNGRRT and TTTN, respectively. The combination of three CRISPR proteins expanded the targetable sites to an average of 6.5 bp resolutions.…”

Section: Current Prospective and Future Directionsmentioning

confidence: 99%

“…Genome editing has rapidly spread from a few model organisms such as roundworm, Drosophila, Arabidopsis and mice, to nearly any microbes, plants and animals, as well as human embryos (Liang et al, 2017;Ma et al, 2017). Since the invention of ZFN technology, B. mori has fortunately joined the first echelon of genome-edited organisms.…”

Section: B Mori Genome Editing Advances and Technological Progressmentioning

confidence: 99%

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Genome editing in Bombyx mori: New opportunities for silkworm functional genomics and the sericulture industry

Smagghe

Xia

2018

Insect Science

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In recent years, research in life sciences has been remarkably revolutionized owing to the establishment, development and application of genome editing technologies. Genome editing has not only accelerated fundamental research but has also shown promising applications in agricultural breeding and therapy. In particular, the clustered, regularly interspaced, short palindromic repeat (CRISPR) technology has become an indispensable tool in molecular biology owing to its high efficacy and simplicity. Genome editing tools have also been established in silkworm (Bombyx mori), a model organism of Lepidoptera insects with high economic importance. This has remarkably improved the level and scope of silkworm research and could reveal new mechanisms or targets in basic entomology and pest management studies. In this review, we summarize the progress and potential of genome editing in silkworm and its applications in functional genomic studies for generating novel genetic materials.

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Section: Current Prospective and Future Directionsmentioning

confidence: 99%

Section: B Mori Genome Editing Advances and Technological Progressmentioning

confidence: 99%

Genome editing in Bombyx mori: New opportunities for silkworm functional genomics and the sericulture industry

Smagghe

Xia

2018

Insect Science

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“…To determine the efficiency of three delivery methods, we choose BmBlos2 as a target gene, because editing activities can be easily calculated by observing the mosaic oily phenotype. A gRNA (gRNA‐1) targeting the second exon of BmBlos2 was designed and cloned into a vector driven by T7 and BmHsp70 promoters (Ma et al ., ). Moreover, gRNA‐1 was transcribed into RNA using an in vitro transcription kit (MEGAscript, Invitogen).…”

mentioning

confidence: 97%

“…It is generally recognized that off‐target frequency is related to the activity of Cas9/gRNA and exposure time of genomic DNA to Cas9/gRNA, and that limiting Cas9 expression to a narrow time window would reduce the off‐target effect. Previously, Cas9/gRNAs were transfected or injected into cells or embryos in the form of either plasmid DNA or in vitro transcribed RNA (Wang et al ., ; Ma et al ., , ). Cas9 RNPs delivery method, which is a DNA‐free process that shows the highest on‐target activity, is a preferred strategy.…”

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confidence: 99%

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Enhanced Bombyx genome editing via Cas9 ribonucleoprotein injection

Lee

Zhang

et al. 2018

Insect Science

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Cas9 versus Cas12a/Cpf1: Structure–function comparisons and implications for genome editing

2018

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Cas9 and Cas12a are multidomain CRISPR-associated nucleases that can be programmed with a guide RNA to bind and cleave complementary DNA targets. The guide RNA sequence can be varied, making these effector enzymes versatile tools for genome editing and gene regulation applications. While Cas9 is currently the best-characterized and most widely used nuclease for such purposes, Cas12a (previously named Cpf1) has recently emerged as an alternative for Cas9. Cas9 and Cas12a have distinct evolutionary origins and exhibit different structural architectures, resulting in distinct molecular mechanisms. Here we compare the structural and mechanistic features that distinguish Cas9 and Cas12a, and describe how these features modulate their activity. We discuss implications for genome editing, and how they may influence the choice of Cas9 or Cas12a for specific applications. Finally, we review recent studies in which Cas12a has been utilized as a genome editing tool. This article is categorized under: RNA Interactions with Proteins and Other Molecules > Protein-RNA Interactions: Functional Implications Regulatory RNAs/RNAi/Riboswitches > Biogenesis of Effector Small RNAs RNA Interactions with Proteins and Other Molecules > RNA-Protein Complexes.

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An integrated CRISPR Bombyx mori genome editing system with improved efficiency and expanded target sites

Cited by 34 publications

References 35 publications

Genome editing in Bombyx mori: New opportunities for silkworm functional genomics and the sericulture industry

Genome editing in Bombyx mori: New opportunities for silkworm functional genomics and the sericulture industry

Enhanced Bombyx genome editing via Cas9 ribonucleoprotein injection

Cas9 versus Cas12a/Cpf1: Structure–function comparisons and implications for genome editing

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