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Controlled experiment Neocaridina identity evidence

Microinjection-based CRISPR/Cas9 mutagenesis in the decapoda crustaceans Neocaridina heteropoda and Eriocheir sinensis

Li et al., 2022. Journal of Experimental Biology 225(6): jeb243702.

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This record is a claim boundary, not a quality badge. Evidence class and species relationship describe what was studied. They do not make the result universal, complete or automatically applicable to a home aquarium.

Source taxon as published
Animals sold as Neocaridina heteropoda from a local aquatic market and cultured in the laboratory for more than six months; no voucher, morphology or molecular host-identification method reported
Environment
One-cell embryos removed from females four hours after spawning, placed on wet filter paper with the upper surface exposed to air, injected and irrigated three times daily with sterilized freshwater through in vitro development
Life stages
one-cell embryo, gastrula, nauplius, flea larva
Reviewed
2026-08-12

What was studied

Study shape: A tracer pilot compared 1000 ng/uL dextran with 0.05 percent Phenol Red. For editing, three Nh-scarlet sgRNAs were designed using a partial Neocaridina sequence and the Penaeus vannamei genome as reference. Cas9 at 300 ng/uL and sgRNA at 200 ng/uL were delivered at about 0.8 nL per embryo; individual sgRNAs were checked at gastrula and all three were co-injected for the flea-larva eye-phenotype screen.

Experimental unit: Individual embryo for injection and phenotype screening. Twenty-eight injected embryos survived to the reported Neocaridina screen, but the initial number injected, maternal-clutch contributions, allocation, complete control structure and tracer-table denominator or replicate basis were not reported.

Endpoints: embryo survival by developmental stage, Nh-scarlet expression by developmental stage, visible eye pigmentation and shape, target-region PCR and sequencing.

Claim boundary

What it can support: The method delivered CRISPR/Cas9 material into ex vivo Neocaridina embryos and recovered target-region insertions, replacements or deletions with altered eye phenotypes in three of 28 surviving injected embryos: two with altered eye shape and one with an absent eye.

What it cannot support: The authors called 3 of 28 surviving injected embryos a 10 percent gene-editing efficiency; the denominator excludes embryos that did not survive, and the initial injected total was not reported. Screening was based on abnormal eyes, so phenotype-normal survivors were not shown to have been systematically genotyped. Some embryos contained more than one edited sequence, consistent with mosaicism. Off-target effects were not assessed, one sgRNA lacked detectable in vitro digestion, and exact family contributions, randomization, blinding and complete negative-control comparisons were not reported. The study ended at early development and did not test adult survival, health, body colour, fertility, germline transmission, F1 inheritance, stable-line creation, commercial grade, named-trade-line identity or a consumer genetic test. The invasive embryo method is a functional-genomics protocol, not a hobby breeding procedure.

Publication status audit

Matched in dated Crossref query

The canonical DOI appeared in the dated broad query and carried no registered update in that response. This is a metadata observation, not proof that the work has never changed.

Metadata snapshot reviewed 2026-08-12. Inspect the method, unmatched queue and limitations.

Correction impact

If this source boundary changes, these are the known downstream consumers. The list distinguishes scientific re-review from generated parity and historical follow-up.

Editorial review required (8)

This curated wording or synthesis must be reconsidered when the source boundary changes. A passing generator test cannot decide the new scientific meaning.

  • anatomy decision card Tank-side decision card
    The card translates this guide into an observation, first action, next record and stopping boundary, so all four require editorial review when the source changes.
  • breeding decision card Tank-side decision card
    The card translates this guide into an observation, first action, next record and stopping boundary, so all four require editorial review when the source changes.
  • colours decision card Tank-side decision card
    The card translates this guide into an observation, first action, next record and stopping boundary, so all four require editorial review when the source changes.
  • genetics decision card Tank-side decision card
    The card translates this guide into an observation, first action, next record and stopping boundary, so all four require editorial review when the source changes.
  • anatomy practical guide Practical guide
    The guide uses this evidence record and its practical wording must remain inside the revised source boundary.
  • breeding practical guide Practical guide
    The guide uses this evidence record and its practical wording must remain inside the revised source boundary.
  • colours practical guide Practical guide
    The guide uses this evidence record and its practical wording must remain inside the revised source boundary.
  • genetics practical guide Practical guide
    The guide uses this evidence record and its practical wording must remain inside the revised source boundary.

Generated from governing registry (19)

This surface derives from a governing registry and should update with it, but release tests must still prove parity and routing.

Historical record needs follow-up decision (1)

Do not silently rewrite history. Decide whether the prior change record remains accurate and publish a new correction or scope note when needed.

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How this record is classified

Controlled experiment: An assigned treatment or controlled comparison. The treatment, unit, duration and endpoint still limit the claim.

Neocaridina identity evidence: The source addresses identity, relationships or genetics within the Neocaridina complex.

Topics: Genetics, Colour, Anatomy, Breeding

Critically appraise this source without a score

These questions fit this record's controlled experiment role. They are prompts, not automatic judgments. Open the original source to answer them.

  1. Identity and provenance: How were the organism, population, stock and life stage identified and sourced? A published name or seller label can hide a taxonomic, population or life-stage transfer.
  2. Independent unit: What unit was independently assigned or sampled, and how many units support each comparison? Animal counts do not create independent replication when animals share a vessel, site, family or treatment history.
  3. Comparison and context: What was the actual comparator, allocation process, environment, duration and material condition? A result has meaning only against the comparison and conditions that produced it.
  4. Endpoint and measurement: Which endpoint was measured, with what method, unit, timing, resolution and decision rule? One endpoint cannot silently become survival, welfare, diagnosis, reproduction or long-term population performance.
  5. Denominators and missingness: Are starting counts, exclusions, losses, missing observations and analysis denominators reconciled? Unreported or changing denominators can alter the apparent direction, precision and applicability of a result.
  6. Transfer boundary: What is the nearest tempting aquarium claim that this source design cannot establish? Direct evidence can still be narrow, and adjacent evidence can be useful only while the inference remains visible.
  7. Treatment assignment: Were independent units assigned to treatments, and were baseline conditions comparable? Unclear allocation or baseline imbalance can confound the treatment comparison.
  8. Replication: Were treatment vessels or other exposure units independently replicated rather than subsampled? Repeated animals or assays inside one exposure unit do not replicate the environmental treatment.
  9. Exposure verification: Was the treatment, dose, feed intake or environmental exposure measured and maintained as described? Nominal treatment labels may not equal the exposure animals actually received.
  10. Analysis: Did the statistical model match the unit, repeated measures, multiplicity and missing outcomes? A precise p-value cannot repair a mismatched unit or unaccounted comparison structure.

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