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UgRNA/Cas9 non-specific guide RNA system

Candidate: toolkit itemType: construct pattern1 source documents6 linked claims
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Aliases

UgRNA/Cas9

Extracted Explainers

What the tool is doing

UgRNA/Cas9 is used for multi-site, diversified editing of the srfA operon in Bacillus pumilus LG3145. In this paper it directs replacement of adenylation-domain Stachelhaus-code residues to alter surfactin products.

Source 1DOIPubMed

Resources required

The system requires UgRNAs designed against four conserved motifs around the SrfA active pocket and Cas9-mediated editing in the Bacillus pumilus LG3145 strain.

Source 1DOIPubMed

What problem it solves

It addresses the challenge of precise and efficient engineering of NRPS adenylation domains to generate diverse surfactin variants with altered bioactivity.

Source 1DOIPubMed

What it does not solve

The abstract does not show that the system broadly solves editing across other NRPS systems or that all designed amino-acid substitutions were recovered as products.

Source 1DOIPubMed

Alternatives

The abstract contrasts this approach with the general field challenge of engineering NRPS adenylation domains, but does not name a specific alternative editing platform in the source text.

Source 1DOIPubMed

Evidence Snippets

Herein, we applied a non-specific guide RNA system (UgRNA/Cas9) for multi-site and diversified editing of the srfA operon in the high-secreting strain Bacillus pumilus LG3145.
Evidence 1Source 1DOIPubMedprovenance

Supporting Sources

Linked Claims

Claim 1applicationsupports2026Source 1DOIPubMed

UgRNA/Cas9 was applied for multi-site and diversified editing of the srfA operon in Bacillus pumilus LG3145.

Quoted textsource-backed
Herein, we applied a non-specific guide RNA system (UgRNA/Cas9) for multi-site and diversified editing of the srfA operon in the high-secreting strain Bacillus pumilus LG3145.
Claim 2design strategysupports2026Source 1DOIPubMed

UgRNAs targeting four conserved motifs around the SrfA active pocket were designed to direct Cas9-mediated replacement of Stachelhaus codes with codons for 28 different amino acids.

Quoted textsource-backed
UgRNAs targeting four conserved motifs around the SrfA active pocket were designed to direct Cas9-mediated replacement of Stachelhaus codes with codons for 28 different amino acids.
Claim 3engineering outcomesupports2026Source 1DOIPubMed

The editing strategy generated engineered strain WA1 with a module 2 Val-to-Phe mutation that produces [Ser2]surfactin.

Quoted textsource-backed
This strategy generated two engineered strains: WA1, which carries a Val→Phe mutation in module 2 and produces [Ser2]surfactin
Claim 4engineering outcomesupports2026Source 1DOIPubMed

The editing strategy generated engineered strain WA2 with three mutations across modules 3 and 6 that yields [Lys3,6]surfactin.

Quoted textsource-backed
WA2, which harbors three mutations (Thr→Ser, Glu→Lys, Asn→Gly) across modules 3 and 6, yielding [Lys3,6]surfactin.
Claim 5functional effectsupports2026Source 1DOIPubMed

The engineered surfactin variants displayed distinct antifungal profiles compared with the parent strain LG3145.

Quoted textsource-backed
The two variants displayed distinct antifungal profiles: WA1 exhibited strong activity against Ustilaginoidea virens, whereas WA2 inhibited both Pyricularia oryzae and U. virens. In contrast, the parent strain LG3145 was only effective against Rhizoctonia solani.
Claim 6potentialsupports2026Source 1DOIPubMed

UgRNA/Cas9-driven editing of NRPS domains can expand the structural and functional diversity of surfactins.

Quoted textsource-backed
This work demonstrates the potential of UgRNA/Cas9-driven editing of NRPS domains to expand structural and functional diversity of surfactins.