Ingestion of single guide RNAs induces gene overexpression and extends lifespan in Caenorhabditis elegans via CRISPR activation

Fischer, Fabian; Benner, Christoph; Goyala, Anita; Grigolon, Giovanna; Vitiello, Davide; Wu, Jia Yee; Zarse, Kim; Ewald, Collin Y.; Ristow, Michael

doi:10.1016/j.jbc.2022.102085

Cited by 6 publications

(2 citation statements)

References 66 publications

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“…Despite having less internalization in terms of localization than naked dsRNA, they discovered that chitosan/dsRNA polyplex nanoparticles were substantially more efficient at knocking down genes at the whole-body concentration. More recently, CRISPR/Cas9 mediated gene activation is achieved by delivering sgRNA using a bacterium feeding system, and a recent work recently used electroporation to introduce nucleic acids into worms at a population level (Khodakova, 2021;Luo, 2022;Fischer, 2022). Similar to all these approaches, our new strategy sought to quickly and consistently supply nucleic acids to the multicellular organisms at the population level, and nucleic acid was successfully delivered via our polymer-mediated nucleic acid delivery technique.…”

Section: Discussionmentioning

confidence: 99%

Development of Polymer/DNA Polyplexes System for Nucleic Acid Delivery to the Multicellular OrganismC. elegans

Yenisert

Bayram

Koseoglu

et al. 2022

Preprint

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Gene therapy studies have been of great importance in the elimination of genetic diseases, and the capability of the CRISPR/Cas9 genome editing technique to correct genetic defects has shown great promise. As DNA-based Cas9 nuclease delivery is preferable because of its low cost and higher stability, effective vector-based CRISPR/Cas9 administration is urgently needed. Here, we used the multicellular organism Caenorhabditis elegans to optimize the polymer-mediated DNA delivery system to generate mutants with CRISPR/Cas9. Toward this end, the cationically quaternized polymer of POEGMA-b-P4VP (POEGMA-b-QP4VP) as a carrier of CRISPR/Cas9 components was first synthesized, followed by the formation of plasmid DNA-polymer complex called polyplexes. 1H NMR, Zeta-Sizer, Scanning Electron Microscopy (SEM) analysis, and gel retardation experiments confirmed the polyplexes formation, including pRF4 (Roller) and sgRNA dpy-10, which were then incubated with C. elegans. The polymer-mediated delivery system facilitated the generation of transgenic Roller animals and heritable Dumpy mutants with CRISPR/Cas9. Our study for the first time demonstrated optimized administration of CRISPR/Cas 9 components to C. elegans.

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Section: Discussionmentioning

confidence: 99%

Development of Polymer/DNA Polyplexes System for Nucleic Acid Delivery to the Multicellular OrganismC. elegans

Yenisert

Bayram

Koseoglu

et al. 2022

Preprint

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“…This is mainly because genome editing relies on a slow, multi-step microinjection process. Given that a recent study demonstrated the successful delivery of sgRNA [21], feeding-based CRISPR may be an alternative method for genome editing.…”

Section: Elegans For Crispr-cas Genome Editingmentioning

confidence: 99%

A Decade of CRISPR-Cas Gnome Editing in C. elegans

Kim

Hong

Chen

2022

IJMS

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CRISPR-Cas allows us to introduce desired genome editing, including mutations, epitopes, and deletions, with unprecedented efficiency. The development of CRISPR-Cas has progressed to such an extent that it is now applicable in various fields, with the help of model organisms. C. elegans is one of the pioneering animals in which numerous CRISPR-Cas strategies have been rapidly established over the past decade. Ironically, the emergence of numerous methods makes the choice of the correct method difficult. Choosing an appropriate selection or screening approach is the first step in planning a genome modification. This report summarizes the key features and applications of CRISPR-Cas methods using C. elegans, illustrating key strategies. Our overview of significant advances in CRISPR-Cas will help readers understand the current advances in genome editing and navigate various methods of CRISPR-Cas genome editing.

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Caenorhabditis elegans models of tauopathy

Abd El Hady,

Mohsen

2025

Translational Models of Parkinson' S Disease and Related Movement Disorders

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Ingestion of single guide RNAs induces gene overexpression and extends lifespan in Caenorhabditis elegans via CRISPR activation

Cited by 6 publications

References 66 publications

Development of Polymer/DNA Polyplexes System for Nucleic Acid Delivery to the Multicellular OrganismC. elegans

Development of Polymer/DNA Polyplexes System for Nucleic Acid Delivery to the Multicellular OrganismC. elegans

A Decade of CRISPR-Cas Gnome Editing in C. elegans

Caenorhabditis elegans models of tauopathy

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