Abstract
Mutagenesis-based screens in mice are a powerful discovery platform to identify novel genes or gene functions associated with disease phenotypes. An N-ethyl-N-nitrosourea (ENU) mutagenesis screen induces single nucleotide variants randomly in the mouse genome. Subsequent phenotyping of mutant and wildtype mice enables the identification of mutated pathways resulting in phenotypes associated with a particular ENU lesion. This unbiased approach to gene discovery conducts the phenotyping with no prior knowledge of the functional mutations. Before the advent of affordable next generation sequencing (NGS), ENU variant identification was a limiting step in gene characterization, akin to ‘finding a needle in a haystack’. The emergence of a reliable reference genome alongside advances in NGS has propelled ENU mutation discovery from an arduous, time-consuming exercise to an effective and rapid form of mutation discovery. This has permitted large mouse facilities worldwide to use ENU for novel mutation discovery in a high-throughput manner, helping to accelerate basic science at the mechanistic level. Here, we describe three different strategies used to identify ENU variants from NGS data and some of the subsequent steps for mutation characterisation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 486-500 |
| Number of pages | 15 |
| Journal | Mammalian Genome |
| Volume | 26 |
| Issue number | 9-10 |
| DOIs | |
| State | Published - Oct 1 2015 |
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ASJC Scopus subject areas
- Genetics
Cite this
Current strategies for mutation detection in phenotype-driven screens utilising next generation sequencing. / Simon, Michelle M.; Moresco, Eva Marie Y; Bull, Katherine R.; Kumar, Saumya; Mallon, Ann Marie; Beutler, Bruce; Potter, Paul K.
In: Mammalian Genome, Vol. 26, No. 9-10, 01.10.2015, p. 486-500.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Current strategies for mutation detection in phenotype-driven screens utilising next generation sequencing
AU - Simon, Michelle M.
AU - Moresco, Eva Marie Y
AU - Bull, Katherine R.
AU - Kumar, Saumya
AU - Mallon, Ann Marie
AU - Beutler, Bruce
AU - Potter, Paul K.
PY - 2015/10/1
Y1 - 2015/10/1
N2 - Mutagenesis-based screens in mice are a powerful discovery platform to identify novel genes or gene functions associated with disease phenotypes. An N-ethyl-N-nitrosourea (ENU) mutagenesis screen induces single nucleotide variants randomly in the mouse genome. Subsequent phenotyping of mutant and wildtype mice enables the identification of mutated pathways resulting in phenotypes associated with a particular ENU lesion. This unbiased approach to gene discovery conducts the phenotyping with no prior knowledge of the functional mutations. Before the advent of affordable next generation sequencing (NGS), ENU variant identification was a limiting step in gene characterization, akin to ‘finding a needle in a haystack’. The emergence of a reliable reference genome alongside advances in NGS has propelled ENU mutation discovery from an arduous, time-consuming exercise to an effective and rapid form of mutation discovery. This has permitted large mouse facilities worldwide to use ENU for novel mutation discovery in a high-throughput manner, helping to accelerate basic science at the mechanistic level. Here, we describe three different strategies used to identify ENU variants from NGS data and some of the subsequent steps for mutation characterisation.
AB - Mutagenesis-based screens in mice are a powerful discovery platform to identify novel genes or gene functions associated with disease phenotypes. An N-ethyl-N-nitrosourea (ENU) mutagenesis screen induces single nucleotide variants randomly in the mouse genome. Subsequent phenotyping of mutant and wildtype mice enables the identification of mutated pathways resulting in phenotypes associated with a particular ENU lesion. This unbiased approach to gene discovery conducts the phenotyping with no prior knowledge of the functional mutations. Before the advent of affordable next generation sequencing (NGS), ENU variant identification was a limiting step in gene characterization, akin to ‘finding a needle in a haystack’. The emergence of a reliable reference genome alongside advances in NGS has propelled ENU mutation discovery from an arduous, time-consuming exercise to an effective and rapid form of mutation discovery. This has permitted large mouse facilities worldwide to use ENU for novel mutation discovery in a high-throughput manner, helping to accelerate basic science at the mechanistic level. Here, we describe three different strategies used to identify ENU variants from NGS data and some of the subsequent steps for mutation characterisation.
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U2 - 10.1007/s00335-015-9603-x
DO - 10.1007/s00335-015-9603-x
M3 - Article
C2 - 26449678
AN - SCOPUS:84943790784
VL - 26
SP - 486
EP - 500
JO - Mammalian Genome
JF - Mammalian Genome
SN - 0938-8990
IS - 9-10
ER -