published-canonicalmechanismmaha-epistemic/1.0

Guide RNA and PAM recognition

The targeting mechanism that combines guide–DNA complementarity with protospacer-adjacent-motif recognition by Cas9. This candidate preserves the experimental unit—editor or circuit, target, cell system, delivery, protocol, assay, comparator, and observation window—before any broader inference is considered.

Bounded definition

The targeting mechanism that combines guide–DNA complementarity with protospacer-adjacent-motif recognition by Cas9.

What the cited work establishes

The study reconstitutes RNA-programmed Cas9 cleavage in vitro and shows that a designed single-guide RNA can direct sequence-specific DNA cleavage adjacent to an appropriate motif.

The constructs, biological systems, protocols, assays, datasets, and comparisons reported in A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.

Claims: urn:maha:claim:guide-rna-and-pam-recognition

What remains a separate question

Sequence recognition rules do not by themselves predict chromatin access, cellular activity, or every off-target interaction.

An in-vitro nuclease mechanism does not establish editing efficiency, specificity, delivery, safety, or phenotype in cells or organisms.

Connected domain graph

Typed dependencies preserve publication state.

Only independently canonical records receive public links and relation statements. Draft graph topology remains private.

mechanistic dependencycanonical

CRISPR-Cas9 nuclease editing

inbound connection · concept

Target recognition depends on guide complementarity and a compatible adjacent motif.

mechanistic dependencycanonical

CRISPR-Cas9 nuclease editing

outbound connection · concept

Guide and motif recognition direct the nuclease mechanism.

Claim ledger

Every proposition keeps its own evidence state.

empirical-claimsingle-study

The cited experiments show that guide sequence and adjacent motif requirements condition Cas9 binding and cleavage in the reconstructed system.

Scope
The constructs, biological systems, protocols, assays, datasets, and comparisons reported in A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.
Boundary
Sequence recognition rules do not by themselves predict chromatin access, cellular activity, or every off-target interaction.
Uncertainty
There is no universal effect estimate for this method; numerical results remain attached to the source experiment, biological system, assay, and analysis choices.
Replication
This candidate records one bounded source package. Independent replications and contradictory results must be compiled separately before evidence maturity is upgraded.

Primary sources

Citation, locator, rights, and boundary travel together.

  1. Source 1 · Science

    A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity

    Martin Jinek, Krzysztof Chylinski, Ines Fonfara, Michael Hauer, Jennifer A. Doudna, Emmanuelle Charpentier

    Exact locator
    Abstract; Figures 1–5; supplementary materials describing guide design and cleavage assays.
    Establishes
    The study reconstitutes RNA-programmed Cas9 cleavage in vitro and shows that a designed single-guide RNA can direct sequence-specific DNA cleavage adjacent to an appropriate motif.
    Boundary
    An in-vitro nuclease mechanism does not establish editing efficiency, specificity, delivery, safety, or phenotype in cells or organisms.
    Rights basis
    citation with paraphrase · Maha paraphrases the source-level result and links to the version of record; no article passage is reproduced.