AGROBEST: A Highly Efficient Agrobacterium-Mediated Transient Expression System in Arabidopsis Seedlings

Wu, Hung‐Yi; Lai, Erh‐Min

doi:10.1007/978-1-0716-1791-5_7

Plant Synthetic Biology

2022

DOI: 10.1007/978-1-0716-1791-5_7

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AGROBEST: A Highly Efficient Agrobacterium-Mediated Transient Expression System in Arabidopsis Seedlings

Hung‐Yi Wu

Erh‐Min Lai

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Dual activities of an X-family DNA polymerase regulate CRISPR-induced insertional mutagenesis across species

Weiss,

Kumar,

Chen

et al. 2024

Nat Commun

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The canonical non-homologous end joining (c-NHEJ) repair pathway, generally viewed as stochastic, has recently been shown to produce predictable outcomes in CRISPR-Cas9 mutagenesis. This predictability, mainly in 1-bp insertions and small deletions, has led to the development of in-silico prediction programs for various animal species. However, the predictability of CRISPR-induced mutation profiles across species remained elusive. Comparing CRISPR-Cas9 repair outcomes between human and plant species reveals significant differences in 1-bp insertion profiles. The high predictability observed in human cells links to the template-dependent activity of human Polλ. Yet plant Polλ exhibits dual activities, generating 1-bp insertions through both templated and non-templated manners. Polλ knockout in plants leads to deletion-only mutations, while its overexpression enhances 1-bp insertion rates. Two conserved motifs are identified to modulate plant Polλ‘s dual activities. These findings unveil the mechanism behind species-specific CRISPR-Cas9-induced insertion profiles and offer strategies for predictable, precise genome editing through c-NHEJ.

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Dual activities of an X-family DNA polymerase regulate CRISPR-induced insertional mutagenesis across species

Weiss,

Kumar,

Chen

et al. 2024

Nat Commun

View full text Add to dashboard Cite

show abstract

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AGROBEST: A Highly Efficient Agrobacterium-Mediated Transient Expression System in Arabidopsis Seedlings

Cited by 1 publication

References 35 publications

Dual activities of an X-family DNA polymerase regulate CRISPR-induced insertional mutagenesis across species

Dual activities of an X-family DNA polymerase regulate CRISPR-induced insertional mutagenesis across species

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