ZMADM (zebrafish mosaic analysis with double markers) for single-cell gene knockout and dual-lineage tracing.

ABSTRACT: As a vertebrate model organism, zebrafish has many unique advantages in developmental studies, regenerative biology, and disease modeling. However, tissue-specific gene knockout in zebrafish is challenging due to technical difficulties in making floxed alleles. Even when successful, tissue-level knockout can affect too many cells, making it difficult to distinguish cell autonomous from noncell autonomous gene function. Here, we present a genetic system termed zebrafish mosaic analysis with double markers (zMADM). Through Cre/loxP-mediated interchromosomal mitotic recombination of two reciprocally chimeric fluorescent genes, zMADM generates sporadic (<0.5%), GFP⁺ mutant cells along with RFP⁺ sibling wild-type cells, enabling phenotypic analysis at single-cell resolutio

SUBMITTER: Xu B 

PROVIDER: S-EPMC8892518 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature