ArticleNature protocols2018
Application and optimization of CRISPR-Cas9-mediated genome engineering in axolotl (Ambystoma mexicanum).
Article in Nature protocols, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 28 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
28 citing papers in PubMed.
- Tissue composition shapes differential skeletal integration strategies during axolotl limb regeneration.Nature communications · 2026Article
- Protocol for non-invasive genotyping of axolotls using skin swabbing.STAR protocols · 2026Article
- Divergent stem cell mechanisms govern the primary body axis and appendage regeneration in the axolotl.Science advances · 2026Article
- Ependymoglial cells are critical for cortex regeneration in axolotls.Nature communications · 2026Article
- Optimized toolkit for the manipulation of immortalized axolotl fibroblasts.Methods (San Diego, Calif.) · 2025Article
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- Revealing the biological features of the axolotl pancreas as a new research model.Frontiers in cell and developmental biology · 2025Article
- Autonomous Feedback-Driven Engineered DNAzyme-Coated Trojan Horse-like Nanocapsules for On-Demand CRISPR/Cas9 Delivery.ACS nano · 2024Article
- A perspective on neuroethology: what the past teaches us about the future of neuroethology.Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology · 2024Review
- The salamander limb: a perfect model to understand imperfect integration during skeletal regeneration.Biology open · 2024Review
- Baculovirus Production and Infection in Axolotls.Methods in molecular biology (Clifton, N.J.) · 2023Article
- The salamander blastema within the broader context of metazoan regeneration.Frontiers in cell and developmental biology · 2023Review
- A Practical Guide for CRISPR-Cas9-Induced Mutations in Axolotls.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Rebuilding limbs, one cell at a time.Developmental dynamics : an official publication of the American Association of Anatomists · 2022Article
- The specialist in regeneration-the Axolotl-a suitable model to study bone healing?NPJ Regenerative medicine · 2022Article
- The use of transgenics in the laboratory axolotl.Developmental dynamics : an official publication of the American Association of Anatomists · 2022Review
- Gene and transgenics nomenclature for the laboratory axolotl-Ambystoma mexicanum.Developmental dynamics : an official publication of the American Association of Anatomists · 2022Article
- Tig1 regulates proximo-distal identity during salamander limb regeneration.Nature communications · 2022Article
- The Axolotl's journey to the modern molecular era.Current topics in developmental biology · 2022Article
Corrections and comments
- Erratum issued
Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Genomic manipulation is essential to the use of model organisms to understand development, regeneration and adult physiology. The axolotl (Ambystoma mexicanum), a type of salamander, exhibits an unparalleled regenerative capability in a spectrum of complex tissues and organs, and therefore serves as a powerful animal model for dissecting mechanisms of regeneration. We describe here an optimized stepwise protocol to create genetically modified axolotls using the CRISPR-Cas9 system. The protocol, which takes 7-8 weeks to complete, describes generation of targeted gene knockouts and knock-ins and includes site-specific integration of large targeting constructs. The direct use of purified CAS9-NLS (CAS9 containing a C-terminal nuclear localization signal) protein allows the prompt formation of guide RNA (gRNA)-CAS9-NLS ribonucleoprotein (RNP) complexes, which accelerates the creation of double-strand breaks (DSBs) at targeted genomic loci in single-cell-stage axolotl eggs. With this protocol, a substantial number of F
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