Creating a "hopeful monster": mouse forward genetic screens - PubMed (original) (raw)

Creating a "hopeful monster": mouse forward genetic screens

Vanessa L Horner et al. Methods Mol Biol. 2011.

Abstract

One of the most straightforward approaches to making novel biological discoveries is the forward genetic screen. The time is ripe for forward genetic screens in the mouse since the mouse genome is sequenced, but the function of many of the genes remains unknown. Today, with careful planning, such screens are within the reach of even small individual labs. In this chapter we first discuss the types of screens in existence, as well as how to design a screen to recover mutations that are relevant to the interests of a lab. We then describe how to create mutations using the chemical N-ethyl-N-nitrosourea (ENU), including a detailed injection protocol. Next, we outline breeding schemes to establish mutant lines for each type of screen. Finally, we explain how to map mutations using recombination and how to ensure that a particular mutation causes a phenotype. Our goal is to make forward genetics in the mouse accessible to any lab with the desire to do it.

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Figures

Figure 11.1

Figure 11.1

Approximate breeding space required per month in a generic yearlong screen for recessive mutations.

Figure 11.2

Figure 11.2

Classes of genome-wide and region-specific forward genetic screens. Adapted by permission from Macmillan Publishers Ltd: Nat Rev Genet (33), copyright 2005.

Figure 11.3

Figure 11.3

Crossing scheme for dominant (upper grey box) or recessive (lower grey box) mutant alleles. In this and all subsequent figures, chromosomes from the mutagenized black mouse are represented as black bars; chromosomes from the white mouse are represented as white bars. Additionally, in all figures stars represent mutant alleles.

Figure 11.4

Figure 11.4

Crossing scheme for dominant (upper grey box) or recessive (lower grey box) modifier alleles. In this crossing scheme the allele to be modified (in the white mouse) is assumed to be homozygous lethal or sterile. The half-black/half-white chromosome in the second generation indicates that either allele is acceptable in this cross.

Figure 11.5

Figure 11.5

Non-complementation crossing scheme. (A) The allele to be tested (in white mouse) is viable and fertile as a homozygote. (B) The allele to be tested (in white mouse) is lethal or sterile as a homozygote.

Figure 11.6

Figure 11.6

Deletion screen crossing scheme. The chromosome with a gap indicates the region that is deleted. Mutant alleles are recovered in the second generation (grey box).

Figure 11.7

Figure 11.7

Balancer screen crossing scheme. The white bar with double-sided arrows indicates the balanced chromosome. In the first generation, F1 mice are crossed to mice carrying the inversion in trans to a WT chromosome marked with a dominant visible mutation (dotted bar with black circle). Mutant alleles are recovered in the third generation (grey box).

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