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Multiplex CRISPR/Cas screen in regenerating haploid limbs of chimeric Axolotls.
- Source :
-
ELife [Elife] 2020 Jan 28; Vol. 9. Date of Electronic Publication: 2020 Jan 28. - Publication Year :
- 2020
-
Abstract
- Axolotls and other salamanders can regenerate entire limbs after amputation as adults, and much recent effort has sought to identify the molecular programs controlling this process. 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.<br />Competing Interests: LS, GF, CC No competing interests declared<br /> (© 2020, Sanor et al.)
- Subjects :
- Alleles
Ambystoma mexicanum embryology
Animals
Catalase genetics
Fetuin-B genetics
Gene Editing
Gene Expression Regulation, Developmental
Genetic Testing methods
Karyotype
Methyltransferases genetics
Monophenol Monooxygenase genetics
Mutation
Regeneration
Sequence Analysis
Ambystoma mexicanum genetics
Ambystoma mexicanum physiology
CRISPR-Cas Systems
Haploidy
Subjects
Details
- Language :
- English
- ISSN :
- 2050-084X
- Volume :
- 9
- Database :
- MEDLINE
- Journal :
- ELife
- Publication Type :
- Academic Journal
- Accession number :
- 31989926
- Full Text :
- https://doi.org/10.7554/eLife.48511