A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila

Huynh, Nhan; Depner, Noah; Larson, Raegan T.; King-Jones, Kirst

doi:10.1186/s13059-020-02193-y

Cited by 79 publications

(65 citation statements)

References 119 publications

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“…One explanation is that previous reports of Cas13 function in plants and other systems have not included a guide-alone control (e.g. stable transgenic line expressing guide crRNA alone) such as the experiments described for stable transgenic rice 17 , rice protoplasts 15 , and experiments in animal systems 15,44,45 . One experiment did test for guide crRNA-alone effects against TuMV in N. benthamiana, but reported no impact on viral accumulation 16 .…”

Section: Discussionmentioning

confidence: 99%

RNA silencing by CRISPR in plants does not require Cas13

Sharma

Marla

Zheng

et al. 2021

Preprint

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RNA-targeting CRISPR-Cas can provide potential advantages over DNA editing, such as avoiding pleiotropic effects of genome editing, providing precise spatiotemporal regulation and expanded function including anti-viral immunity. Here, we report the use of CRISPR-Cas13 in plants to reduce both viral and endogenous RNA. Unexpectedly, we discovered that crRNA designed to guide Cas13 could, in the absence of the Cas13 protein, cause substantial reduction in RNA levels as well. We demonstrate Cas13-independent guide-induced gene silencing (GIGS) in three plant species, including stable transgenic Arabidopsis. We determined that GIGS utilizes endogenous RNAi machinery despite the fact that crRNA are unlike canonical triggers of RNAi such as miRNA, hairpins or long double-stranded RNA. These results suggest that GIGS offers a novel and flexible approach to RNA reduction with potential benefits over existing technologies for crop improvement. Our results demonstrate that GIGS is active across a range of plant species, evidence similar to recent findings in an insect system, which suggests that GIGS is potentially active across many eukaryotes.

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Section: Discussionmentioning

confidence: 99%

RNA silencing by CRISPR in plants does not require Cas13

Sharma

Marla

Zheng

et al. 2021

Preprint

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“…The recently discovered Cas13 family members are smaller than Cas9 and use only a single short CRISPR RNA to target RNA as their substrate ( Abudayyeh et al, 2016 ; East-Seletsky et al, 2016 ; Konermann et al, 2018 ; Yan et al, 2018b ). A Drosophila -optimized and MTS-tagged variant of the Cas13 family member Cas13d successfully targets transcripts of the OXPHOS genes mt:CoI or mt:CoII , the levels of which it could reduce by four- to fivefold ( Huynh et al, 2020 ). In the future, mitochondrially targeted DNA and RNA nucleases and base editors should facilitate the study of mitochondrial diseases.…”

Section: Mitochondrial Diseasesmentioning

confidence: 99%

Mitochondrial function in development and disease

Rossmann

Dubois

Agarwal

et al. 2021

Disease Models &Amp; Mechanisms

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Mitochondria are organelles with vital functions in almost all eukaryotic cells. Often described as the cellular ‘powerhouses’ due to their essential role in aerobic oxidative phosphorylation, mitochondria perform many other essential functions beyond energy production. As signaling organelles, mitochondria communicate with the nucleus and other organelles to help maintain cellular homeostasis, allow cellular adaptation to diverse stresses, and help steer cell fate decisions during development. Mitochondria have taken center stage in the research of normal and pathological processes, including normal tissue homeostasis and metabolism, neurodegeneration, immunity and infectious diseases. The central role that mitochondria assume within cells is evidenced by the broad impact of mitochondrial diseases, caused by defects in either mitochondrial or nuclear genes encoding for mitochondrial proteins, on different organ systems. In this Review, we will provide the reader with a foundation of the mitochondrial ‘hardware’, the mitochondrion itself, with its specific dynamics, quality control mechanisms and cross-organelle communication, including its roles as a driver of an innate immune response, all with a focus on development, disease and aging. We will further discuss how mitochondrial DNA is inherited, how its mutation affects cell and organismal fitness, and current therapeutic approaches for mitochondrial diseases in both model organisms and humans.

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“…In multiplex RNA editing, pre-crRNA processing ability of Cas13, can be used to target multiple RNA targets by processing multiple crRNA from a single array. Unlike Cas9 and Cas12, which recognize PAM sequences adjacent to a target sequence, Cas13a recognizes a PFS followed by a target sequence [ 105 ]. However, while LshCas13a from Leptotrichia shahii requires a PFS for target identification [ 67 ], LwaCas13a (from Leptotrichia wadei ) and PsPCas13b (from Prevotella spp.)…”

Section: Emerging Crispr/cas Systems and Their Applicationsmentioning

confidence: 99%

An Outlook on Global Regulatory Landscape for Genome-Edited Crops

Ahmad

Munawar

Khan

et al. 2021

IJMS

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The revolutionary technology of CRISPR/Cas systems and their extraordinary potential to address fundamental questions in every field of biological sciences has led to their developers being awarded the 2020 Nobel Prize for Chemistry. In agriculture, CRISPR/Cas systems have accelerated the development of new crop varieties with improved traits—without the need for transgenes. However, the future of this technology depends on a clear and truly global regulatory framework being developed for these crops. Some CRISPR-edited crops are already on the market, and yet countries and regions are still divided over their legal status. CRISPR editing does not require transgenes, making CRISPR crops more socially acceptable than genetically modified crops, but there is vigorous debate over how to regulate these crops and what precautionary measures are required before they appear on the market. This article reviews intended outcomes and risks arising from the site-directed nuclease CRISPR systems used to improve agricultural crop plant genomes. It examines how various CRISPR system components, and potential concerns associated with CRISPR/Cas, may trigger regulatory oversight of CRISPR-edited crops. The article highlights differences and similarities between GMOs and CRISPR-edited crops, and discusses social and ethical concerns. It outlines the regulatory framework for GMO crops, which many countries also apply to CRISPR-edited crops, and the global regulatory landscape for CRISPR-edited crops. The article concludes with future prospects for CRISPR-edited crops and their products.

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A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila

Cited by 79 publications

References 119 publications

RNA silencing by CRISPR in plants does not require Cas13

RNA silencing by CRISPR in plants does not require Cas13

Mitochondrial function in development and disease

An Outlook on Global Regulatory Landscape for Genome-Edited Crops

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