A genome-wide library of MADM mice for single-cell genetic mosaic analysis

Abstract

Highlights d Genome-wide resource for genetic mosaic analysis with double markers in mice d Resource for dissection of cell-autonomous gene function of >96% of the mouse genome d Resource for genome-wide analysis of genomic imprinting phenotypes d MADM chromosomes reveal non-random mitotic sister chromatid segregation in vivo

Genetic mosaic dissection of candidate genes in mice using mosaic analysis with double markers

“…This protocol describes the generation of genetic mosaic mice using mosaic analysis with double markers (MADM) ( Zong et al., 2005 ; Contreras et al., 2021 ). A genetic mosaic individual consists of cells of distinct genotypes, for example harboring homozygous mutant cells next to wild-type cells.…”

“…Thus, mosaicism enables the study of cell-autonomous gene function at single cell resolution. Individual mutant cells can be studied in many ways, for example in lineage tracing, cell competition assays, morphological analysis and disease modelling ( Hippenmeyer et al., 2010 ; Gao et al., 2014 ; Joo et al., 2014 ; Beattie et al., 2017 ; Tian et al., 2020 ; Contreras et al., 2021 ).…”

“…In this protocol, we present an experimental pipeline for the generation of MADM-labeled mosaic tissues and highlight a variety of downstream analysis applications. Our protocol is based on the use of distinct Cre drivers in combination with MADM cassettes targeted to all 19 mouse autosomes ( Contreras et al., 2021 ).…”

“…This protocol highlights major steps for the generation of genetic mosaic tissue and the isolation and processing of respective tissues for downstream histological analysis. For complete details on the use and execution of this protocol, please refer to Contreras et al. (2021) .…”

“…For complete details on the use and execution of this protocol, please refer to Contreras et al. (2021) .…”