Flexible TAM requirement of TnpB enables efficient single-nucleotide editing with expanded targeting scope DOI Creative Commons
Xu Feng,

Ruyi Xu,

Jianglan Liao

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: April 24, 2024

Abstract TnpBs encoded by the IS200/IS605 family transposon are among most abundant prokaryotic proteins from which type V CRISPR-Cas nucleases may have evolved. Since bacterial can be programmed for RNA-guided dsDNA cleavage in presence of a transposon-adjacent motif (TAM), these hold immense promise genome editing. However, activity and targeting specificity TnpB homology-directed gene editing remain unknown. Here we report that thermophilic archaeal enables efficient natural host. Interestingly, has different TAM requirements eliciting cell death facilitating By systematically characterizing variants, reveal recognizes broad range sequences including those do not elicit apparent death. Importantly, shows very high on targets flanked weak TAM. Taking advantage this feature, successfully leverage single-nucleotide with templated repair. The use only facilitates more flexible increased survival, but also greatly expands scopes, strategy is probably applicable to diverse systems.

Language: Английский

Flexible TAM requirement of TnpB enables efficient single-nucleotide editing with expanded targeting scope DOI Creative Commons
Xu Feng,

Ruyi Xu,

Jianglan Liao

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: April 24, 2024

Abstract TnpBs encoded by the IS200/IS605 family transposon are among most abundant prokaryotic proteins from which type V CRISPR-Cas nucleases may have evolved. Since bacterial can be programmed for RNA-guided dsDNA cleavage in presence of a transposon-adjacent motif (TAM), these hold immense promise genome editing. However, activity and targeting specificity TnpB homology-directed gene editing remain unknown. Here we report that thermophilic archaeal enables efficient natural host. Interestingly, has different TAM requirements eliciting cell death facilitating By systematically characterizing variants, reveal recognizes broad range sequences including those do not elicit apparent death. Importantly, shows very high on targets flanked weak TAM. Taking advantage this feature, successfully leverage single-nucleotide with templated repair. The use only facilitates more flexible increased survival, but also greatly expands scopes, strategy is probably applicable to diverse systems.

Language: Английский

Citations

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