Efficient gene-driven germ-line point mutagenesis of C57BL/6J mice

Abstract

Background: Analysis of an allelic series of point mutations in a gene, generated by N-ethyl-N-nitrosourea (ENU) mutagenesis, is a valuable method for discovering the full scope of its biological function. Here we present an efficient gene-driven approach for identifying ENU-induced point mutations in any gene in C57BL/6J mice. The advantage of such an approach is that it allows one to select any gene of interest in the mouse genome and to go directly from DNA sequence to mutant mice.

Generation of N-Ethyl-N-nitrosourea (ENU) Diabetes Models in Mice Demonstrates Genotype-specific Action of Glucokinase Activators

“…The dose of ENU used has been reported to produce a mutation rate between 2.6 ϫ 10 Ϫ7 and 1.04 ϫ 10 Ϫ6 mutations/base pair in B6 mice (23)(24)(25)(26)(27). As has been argued previously (28), such a mutation rate would produce multiple mutations in a single mouse, but the mutations would be spaced widely enough that they would sort independently during meiosis.…”

Deep genome sequencing and variation analysis of 13 inbred mouse strains defines candidate phenotypic alleles, private variation, and homozygous truncating mutations

“…Of these, 2,462 were novel and 2,110 were private to one of the 13 strains in our study. Similarly, 4,620 structural insertions overlapped the coding sequence of 5,076 genes, many of which were either novel (2,557) or private (2,366) to one of the 13 strains strains.…”

“…Genetic mutations (such as SNPs, indels and structural variants) are commonly involved in the dysregulation of normal gene function and frequently the cause of human diseases. Laboratory mouse strains are one of the primary model organisms used to study human disease, and have excelled as a model system in which to identify and characterise the role that genomic variation plays in susceptibility and pathogenesis [1,2]. A multitude of inbred laboratory strains are now commonly employed in the examination of a wide range of human diseases [3].…”

Designing Phenotyping Studies for Genetically Engineered Mice

“…The goal of KOMP and other high-throughput knockout projects is to observe the effect of loss of function of every gene in the mouse genome. Although these effects may be embryonic lethal and often do not reflect the corresponding human condition, this approach provides an essential platform upon which more subtle defects such as ENU point mutagenesis 53 may be assessed.…”