published-canonicalmeasurementmaha-epistemic/1.0

Targeted amplicon sequencing of edit outcomes

Locus-focused sequencing used to quantify intended edits, indels, and selected nominated off-target outcomes. 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

Locus-focused sequencing used to quantify intended edits, indels, and selected nominated off-target outcomes.

What the cited work establishes

The study develops GUIDE-seq, a cell-based double-strand-break capture method, and applies it to specified nucleases and human cell lines.

The constructs, biological systems, protocols, assays, datasets, and comparisons reported in GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases.

Claims: urn:maha:claim:targeted-amplicon-sequencing

What remains a separate question

Amplicon assays are limited by primer design, sampled sites, allele representation, read depth, error correction, and variant-calling thresholds.

GUIDE-seq has assay-specific detection limits and dependencies and does not nominate every possible off-target event in every biological context.

Connected domain graph

Typed dependencies preserve publication state.

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

Claim ledger

Every proposition keeps its own evidence state.

empirical-claimsingle-study

The cited GUIDE-seq workflow uses targeted sequencing and validation to characterize nominated cleavage sites under specified experiments.

Scope
The constructs, biological systems, protocols, assays, datasets, and comparisons reported in GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases.
Boundary
Amplicon assays are limited by primer design, sampled sites, allele representation, read depth, error correction, and variant-calling thresholds.
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 · Nature Biotechnology

    GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases

    Shengdar Q. Tsai, Zongli Zheng, Nhu T. Nguyen, Matthew Liebers, et al.

    Exact locator
    Abstract; Figures 1–6; Online Methods; Supplementary Tables; Sequence Read Archive SRP050338.
    Establishes
    The study develops GUIDE-seq, a cell-based double-strand-break capture method, and applies it to specified nucleases and human cell lines.
    Boundary
    GUIDE-seq has assay-specific detection limits and dependencies and does not nominate every possible off-target event in every biological context.
    Rights basis
    citation with paraphrase · Maha paraphrases the source-level result and links to the version of record; no article passage is reproduced.