Generating Genetically Modified Mice: A Decision Guide

Methods Mol Biol. 2017;1642:1-19. doi: 10.1007/978-1-4939-7169-5_1.

Abstract

The generation of a new genetically modified mouse strain is a big hurdle to take for many researchers. It is often unclear which steps and decisions have to be made prior to obtaining the desired mouse model. This review aims to help researchers by providing a decision guide that answers the essential questions that need to be asked before generating the most suitable genetically modified mouse line in the most optimal timeframe. The review includes the latest technologies in both the stem cell culture and gene editing tools, particularly CRISPR/Cas9, and provides compatibility guidelines for selecting among the different types of genetic modifications that can be introduced in the mouse genome and the various routes for introducing these modifications into the mouse germline.

Keywords: CRISPR/Cas9; Embryonic stem cell; GEMM-ESC; Gene targeting; Genetic engineering; Mouse models; Recombinase; Transgenesis; Transposon; Zygote.

Publication types

  • Review

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • CRISPR-Associated Protein 9
  • CRISPR-Cas Systems
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • DNA Transposable Elements
  • Disease Models, Animal
  • Endonucleases / genetics
  • Endonucleases / metabolism
  • Founder Effect*
  • Gene Editing / methods*
  • Gene Targeting / methods*
  • Genetic Engineering / methods*
  • Genome*
  • Germ-Line Mutation
  • Mice
  • Mice, Transgenic*
  • Mouse Embryonic Stem Cells / cytology
  • Mouse Embryonic Stem Cells / metabolism
  • RNA, Guide / genetics
  • RNA, Guide / metabolism
  • Research Design

Substances

  • Bacterial Proteins
  • DNA Transposable Elements
  • RNA, Guide
  • CRISPR-Associated Protein 9
  • Cas9 protein, Francisella novicida
  • Endonucleases