Detection of Marker-Free Precision Genome Editing and Genetic Variation through the Capture of Genomic Signatures

Billon, Pierre; Nambiar, Tarun S.; Hayward, Samuel B.; Zafra, María Paz; Schatoff, Emma M.; Oshima, Koichi; Dunbar, Andrew; Breinig, Marco; Park, Young C; Ryu, Han Suk; Tschaharganeh, Darjus F.; Levine, Ross L.; Baer, Richard; Ferrando, Adolfo A.; Dow, Lukas E.; Ciccia, Alberto

doi:10.1016/j.celrep.2020.02.068

Cited by 10 publications

(5 citation statements)

References 66 publications

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“…When possible, we recommend testing editing efficiency in cultured cells before proceeding in vivo. While amplicon sequencing produces highquality quantitative data, there are faster and cheaper molecular assays such as dinucleotide signature capture (53) or tracking of indels by decomposition (54).…”

Section: Discussionmentioning

confidence: 99%

Precise genome engineering in Drosophila using prime editing

Bosch

Birchak

Perrimon

2020

Proc. Natl. Acad. Sci. U.S.A.

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Precise genome editing is a valuable tool to study gene function in model organisms. Prime editing, a precise editing system developed in mammalian cells, does not require double-strand breaks or donor DNA and has low off-target effects. Here, we applied prime editing for the model organismDrosophila melanogasterand developed conditions for optimal editing. By expressing prime editing components in cultured cells or somatic cells of transgenic flies, we precisely introduce premature stop codons in three classical visible marker genes,ebony,white, andforked. Furthermore, by restricting editing to germ cells, we demonstrate efficient germ-line transmission of a precise edit inebonyto 36% of progeny. Our results suggest that prime editing is a useful system inDrosophilato study gene function, such as engineering precise point mutations, deletions, or epitope tags.

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Section: Discussionmentioning

confidence: 99%

Precise genome engineering in Drosophila using prime editing

Bosch

Birchak

Perrimon

2020

Proc. Natl. Acad. Sci. U.S.A.

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“…When possible, we recommend testing editing efficiency in S2R+ cells before proceeding in vivo. While amplicon sequencing produces high quality quantitative data, there are faster and cheaper molecular assays such as the Dinucleotide signaTurE CapTure (DTECT) (BILLON et al 2020) or Tracking of Indels by Decomposition (TIDE) (SENTMANAT et al 2018). In summary, we have developed genetic tools to express prime editing components in Drosophila, and optimized conditions for efficient editing in cultured cells and in vivo.…”

Section: Prime Editing In Vivomentioning

confidence: 99%

Precise genome engineering inDrosophilausing prime editing

Bosch

Birchak

Perrimon

2020

Preprint

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Precise genome editing is a valuable tool to study gene function in model organisms. Prime editing, a precise editing system developed in mammalian cells, does not require double strand breaks or donor DNA and has low off-target effects. Here, we applied prime editing for the model organism Drosophila melanogaster and developed conditions for optimal editing. By expressing prime editing components in cultured cells or somatic cells of transgenic flies, we precisely installed premature stop codons in three classical visible marker genes, ebony, white, and forked. Furthermore, by restricting editing to germ cells, we demonstrate efficient germ line transmission of a precise edit in ebony to ~50% of progeny. Our results suggest that prime editing is a useful system in Drosophila to study gene function, such as engineering precise point mutations, deletions, or epitope tags.

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“…By using the PE technology, the base conversion of target sequence in mice was successfully carried out ( Billon et al, 2020 ; Liu et al, 2020 ; Gao et al, 2021 ; Liu et al, 2021a ). At the same time, off-target editing was found.…”

Section: Application Of the Pe Systemmentioning

confidence: 99%

Current advancement in the application of prime editing

Huang

Liu²

2023

Front. Bioeng. Biotechnol.

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Prime editing (PE) is a precise genome manipulation technology based on the “search and replace” approach of the CRISPR-Cas9 system, while it does not require the exogenous donor DNA and the DNA double-strand breaks (DSBs). Comparing the base editing technology, the editing scope of prime editing has been widely expanded. Prime editing has been successfully applied in a variety of plant cells, animal cells and the model microorganism Escherichia coli so far, and it has shown a good application potential in breeding and genomic functional study of animals and plants, disease treatment, and modification of the microbial strains. In this paper, the basic strategies of prime editing are briefly described, and its research progress is summarized and prospected from the application of multiple species. In addition, a variety of optimization strategies for improving its efficiency and specificity of prime editing are outlined.

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Detection of Marker-Free Precision Genome Editing and Genetic Variation through the Capture of Genomic Signatures

Cited by 10 publications

References 66 publications

Precise genome engineering in Drosophila using prime editing

Precise genome engineering in Drosophila using prime editing

Precise genome engineering inDrosophilausing prime editing

Current advancement in the application of prime editing

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