Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize

Jiang, Yuanyuan; Chai, Yiping; Lu, Minhui; Han, Xiuli; Zhang, Qiang; Zhou, Yun; Wang, Xuechen; Chen, Qijun

doi:10.1101/2020.07.06.188896

Cited by 14 publications

(22 citation statements)

References 17 publications

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“…The vectors described in this study together with their annotated sequences are available from Addgene and/or MolecularCloud (GenScript). NGS data are available under BioProject ID PRJNA661716 (https://www.ncbi.nlm.nih.gov/ bioproject/PRJNA661716) [16] and BioProject ID PRJNA661699 (https://www.ncbi.nlm.nih.gov/bioproject/661699) [17].…”

Section: Rice Protoplast Transfection and Analysis Of Prime Editingmentioning

confidence: 99%

Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize

et al. 2020

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Prime editing is a novel and universal CRISPR/Cas-derived precision genome-editing technology that has been recently developed. However, low efficiency of prime editing has been shown in transgenic rice lines. We hypothesize that enhancing pegRNA expression could improve prime-editing efficiency. In this report, we describe two strategies for enhancing pegRNA expression. We construct a prime editing vector harboring two pegRNA variants for W542L and S621I double mutations in ZmALS1 and ZmALS2. Compared with previous reports in rice, we achieve much higher prime-editing efficiency in maize. Our results are inspiring and provide a direction for the optimization of plant prime editors.

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Section: Rice Protoplast Transfection and Analysis Of Prime Editingmentioning

confidence: 99%

Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize

et al. 2020

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“…Whereas prime editing was originally developed in human cells (13), it has been quickly adopted in other organisms including mice (13,15,16) and plants (17)(18)(19)(20)(21)(22)(23)(24)(25). Prime editing has been used to help correct disease mutations (13,26), introduce herbicide-resistant alleles (18,19,21,24), alter plant morphology (21), and model human disease mutations in organoids (16,27).…”

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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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“…Although it is a relatively new technique, prime editing has been used in many different cell lines and organisms including plants, zebrafish and Drosophila [8][9][10][11][12][13][14][15][16][17][18][19][20]. In this study we have created versatile plasmids to perform prime editing PE2, PE3 and PE-nuclease system in mammalian cells.…”

Section: Discussionmentioning

confidence: 99%

Optimized nickase- and nuclease-based prime editing in human and mouse cells

Adikusuma

Lushington

Arudkumar

et al. 2021

Preprint

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Precise genomic modification using prime editing (PE) holds enormous potential for research and clinical applications. Currently, the delivery of PE components to mammalian cell lines requires multiple plasmid vectors. To overcome this limitation, we generated all-in-one prime editing (PEA1) constructs that carry all the components required for PE, along with a selection marker. We tested these constructs (with selection) in HEK293T, K562, HeLa and mouse embryonic stem (ES) cells. We discovered that PE efficiency in HEK293T cells was much higher than previously observed, reaching up to 95% (mean 67%). The efficiency in K562 and HeLa cells, however, remained low. To improve PE efficiency in K562 and HeLa, we generated a nuclease prime editor and tested this system in these cell lines as well as mouse ES cells. PE-nuclease generated intended edits with efficiencies that were similar, and in some cases exceeded, the PE-nickase system. We also show that the nuclease prime editor can generate intended modifications in mouse fetuses with up to 100% efficiency.

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Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize

Cited by 14 publications

References 17 publications

Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize

Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize

Precise genome engineering in Drosophila using prime editing

Optimized nickase- and nuclease-based prime editing in human and mouse cells

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