Genotyping Genome-Edited Founders and Subsequent Generation

Matthew Mackenzie; Alex Fower; Alasdair J Allan; Gemma F Codner; Rosie K Bunton-Stasyshyn; Lydia Teboul

doi:10.1007/978-1-0716-2990-1_4

Genotyping Genome-Edited Founders and Subsequent Generation

Methods Mol Biol. 2023:2631:103-134. doi: 10.1007/978-1-0716-2990-1_4.

Authors

Matthew Mackenzie¹, Alex Fower¹, Alasdair J Allan¹, Gemma F Codner¹, Rosie K Bunton-Stasyshyn², Lydia Teboul³

Affiliations

¹ The Mary Lyon Centre, MRC Harwell, Didcot, Oxon, UK.
² The Mary Lyon Centre, MRC Harwell, Didcot, Oxon, UK. [email protected].
³ The Mary Lyon Centre, MRC Harwell, Didcot, Oxon, UK. [email protected].

PMID: 36995665
DOI: 10.1007/978-1-0716-2990-1_4

Abstract

Targeted nucleases allow the production of many types of genetic mutations directly in the early embryo. However, the outcome of their activity is a repair event of unpredictable nature, and the founder animals that are produced are generally of a mosaic nature. Here, we present the molecular assays and genotyping strategies that will support the screening of the first generation for potential founders and the validation of positive animals in the subsequent generation, according to the type of mutation generated.

Keywords: Allele QC; CRISPR-Cas9; Embryo/zygote engineering; Genetic engineering; Genome editing; Genotyping; Mouse models; Transgenesis.

MeSH terms

Animals
CRISPR-Cas Systems*
Gene Editing*
Genome
Genotype
Mutation

Grants and funding

MC_UP_2201/3/MRC_/Medical Research Council/United Kingdom