Functional annotation of mammalian genomic DNA sequence by chemical mutagenesis: A fine-structure genetic mutation map of a 1- to 2-cM segment of mouse chromosome 7 corresponding to human chromosome 11p14-p15
Abstract:Eleven independent, recessive, N-ethyl-N-nitrosourea-induced mutations that map to a Ϸ1-to 2-cM region of mouse chromosome (Chr) 7 homologous to human Chr 11p14-p15 were recovered from a screen of 1,218 gametes. These mutations were initially identified in a hemizygous state opposite a large p-locus deletion and subsequently were mapped to finer genomic intervals by crosses to a panel of smaller p deletions. The 11 mutations also were classified into seven complementation groups by pairwise crosses. Four compl… Show more
“…We isolated seven recessive lethal mutations yielding one mutant mouse line for every 136 genomes screened. Our results are comparable with the SLT deletion-based ENU mutagenesis screens covering the albino and pink-eyed dilution regions of chromosome 7 that realized mutation recovery rates of 1/147 and 1/111, respectively (Rinchik and Carpenter, 1999;Rinchik et al, 2002).…”
Section: Screening For Essential Genes Within the 19 Mb 24pub Piebaldsupporting
confidence: 91%
“…Recently, regional screens using balancer chromosomes have cataloged hundreds of essential loci on chromosomes 4, 5, and 11 (Hentges et al, 2007;Kile et al, 2003;Nishijima et al, 2003;Wilson et al, 2005). A number of regional screens have also been performed using the deletion complex resources generated by the specific locus test (SLT) (Rinchik and Carpenter, 1999;Rinchik et al, 2002). The SLT has a rich genetic legacy, including establishing ENU as the most potent point mutagen in the mouse and building the nested panels of deletions centered on the seven SLT loci (Davis and Justice, 1998;Hitotsumachi et al, 1985;Russell, 1951;Russell et al, 1979).…”
The piebald deletion complex is a set of overlapping chromosomal deficiencies on distal mouse chromosome 14. We surveyed the functional genetic content of the piebald deletion region in an essential gene mutagenesis screen of 952 genomes to recover seven lethal mutants. The ENU-induced mutations were mapped to define genetic intervals using the piebald deletion panel. Lethal mutations included loci required for establishment of the left-right embryonic axis and a loss-of-function allele of Phr1 resulting in respiratory distress at birth. A functional map of the piebald region integrates experimental genetic data from the deletion panel, mutagenesis screen, and the targeted disruption of specific genes. A comparison of several genomic intervals targeted in regional mutagenesis screens suggests that the piebald region is characterized by a low gene density and high essential gene density with a distinct genomic content and organization that supports complex regulatory interactions and promotes evolutionary stability.
“…We isolated seven recessive lethal mutations yielding one mutant mouse line for every 136 genomes screened. Our results are comparable with the SLT deletion-based ENU mutagenesis screens covering the albino and pink-eyed dilution regions of chromosome 7 that realized mutation recovery rates of 1/147 and 1/111, respectively (Rinchik and Carpenter, 1999;Rinchik et al, 2002).…”
Section: Screening For Essential Genes Within the 19 Mb 24pub Piebaldsupporting
confidence: 91%
“…Recently, regional screens using balancer chromosomes have cataloged hundreds of essential loci on chromosomes 4, 5, and 11 (Hentges et al, 2007;Kile et al, 2003;Nishijima et al, 2003;Wilson et al, 2005). A number of regional screens have also been performed using the deletion complex resources generated by the specific locus test (SLT) (Rinchik and Carpenter, 1999;Rinchik et al, 2002). The SLT has a rich genetic legacy, including establishing ENU as the most potent point mutagen in the mouse and building the nested panels of deletions centered on the seven SLT loci (Davis and Justice, 1998;Hitotsumachi et al, 1985;Russell, 1951;Russell et al, 1979).…”
The piebald deletion complex is a set of overlapping chromosomal deficiencies on distal mouse chromosome 14. We surveyed the functional genetic content of the piebald deletion region in an essential gene mutagenesis screen of 952 genomes to recover seven lethal mutants. The ENU-induced mutations were mapped to define genetic intervals using the piebald deletion panel. Lethal mutations included loci required for establishment of the left-right embryonic axis and a loss-of-function allele of Phr1 resulting in respiratory distress at birth. A functional map of the piebald region integrates experimental genetic data from the deletion panel, mutagenesis screen, and the targeted disruption of specific genes. A comparison of several genomic intervals targeted in regional mutagenesis screens suggests that the piebald region is characterized by a low gene density and high essential gene density with a distinct genomic content and organization that supports complex regulatory interactions and promotes evolutionary stability.
“…2 (41,45,47). In this scheme, coat color alleles of various severity for albino (c) or pink-eyed dilute (p) were used to distinguish between chromosomes contributed by different parents and, thus, to allow identification of progeny carrying new mutations in the interval of interest (43,46). In this manner, embryonic lethal, developmental mutations, such as eed and fit-1, were first recovered (39,44).…”
Section: Chromosome Engineering and Region-specific Screensmentioning
In the mouse, random mutagenesis with N-ethyl-N-nitrosourea (ENU) has been used since the 1970s in forward mutagenesis screens. However, only in the last decade has ENU mutagenesis been harnessed to generate a myriad of new mouse mutations in large-scale genetic screens and focused, smaller efforts. The development of additional genetic tools, such as balancer chromosomes, refinements in genetic mapping strategies, and evolution of specialized assays, has allowed these screens to achieve new levels of sophistication. The impressive productivity of these screens has led to a deluge of mouse mutants that wait to be harnessed. Here the basic large- and small-scale strategies are described, as are the basics of screen design. Finally, and importantly, this review describes the mechanisms by which such mutants may be accessed now and in the future. Thus, this review should serve both as an overview of the power of forward mutagenesis in the mouse and as a resource for those interested in developing their own screens, adding onto existing efforts, or obtaining specific mouse mutants that have already been generated
“…Some of the chemicals and most of the radiations that were used in these experiments produced deletions of various sizes. These deletions, provided they are viable in the heterozygous state, are very precious tools for a detailed analysis of some specific parts of the genome (Rinchik et al 2002).…”
Section: Inducing Mutations With Chemicalsmentioning
The house mouse has been used as a privileged model organism since the early days of genetics, and the numerous experiments made with this small mammal have regularly contributed to enrich our knowledge of mammalian biology and pathology, ranging from embryonic development to metabolic disease, histocompatibility, immunology, behavior, and cancer. Over the past two decades, a number of large-scale integrated and concerted projects have been undertaken that will probably open a new era in the genetics of the species. The sequencing of the genome, which will allow researchers to make comparisons with other mammals and identify regions conserved by evolution, is probably the most important project, but many other initiatives, such as the massive production of point or chromosomal mutations associated with comprehensive and standardized phenotyping of the mutant phenotypes, will help annotation of the ∼25,000 genes packed in the mouse genome. In the same way, and as another consequence of the sequencing, the discovery of many single nucleotide polymorphisms and the development of new tools and resources, like the Collaborative Cross, will contribute to the development of modern quantitative genetics. It is clear that mouse genetics has changed dramatically over the last 10-15 years and its future looks promising.
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