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While targeted mutagenesis approaches like CRISPR\/Cas9 now permit gene-level investigation of these mechanisms, genetic screening in the axolotl requires an extensive commitment of time and space. Previously, we quantified CRISPR\/Cas9-generated mutations in the limbs of mosaic mutant axolotls before and after regeneration and found that the regenerated limb is a highfidelity replicate of the original limb (Flowers et al. 2017). Here, we circumvent aforementioned genetic screening limitations and present methods for a multiplex CRISPR\/Cas9 haploid screen in chimeric axolotls (MuCHaChA), which is a novel platform for haploid genetic screening in animals to identify genes essential for limb regeneration.<\/jats:p>","DOI":"10.7554\/elife.48511","type":"journal-article","created":{"date-parts":[[2020,1,27]],"date-time":"2020-01-27T19:00:13Z","timestamp":1580151613000},"source":"Crossref","is-referenced-by-count":19,"title":["Multiplex CRISPR\/Cas screen in regenerating haploid limbs of chimeric Axolotls"],"prefix":"10.7554","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2756-0457","authenticated-orcid":true,"given":"Lucas D","family":"Sanor","sequence":"first","affiliation":[{"name":"Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, United 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