EUCOMM

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The European Conditional Mouse Mutagenesis Program or EUCOMM is an EU-funded program to generate a library of mutant mouse embryonic stem cells for research purposes. [1] [2]

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Setup

The EUCOMM program is funded by the European Union sixth R&D programme to make a library of mutant mouse embryonic stem cell clones (ES cells), to enable the mouse research community to rapidly create mutant mice with specific mutant genes. The EUCOMM consortium is a member of the International Knockout Mouse Consortium, (an organisation including the EUCOMM, KOMP, NorCOMM and TIGM consortia) which reflects a commitment to share and promote their products and technology with the international research community. [2]

Stem cell creation

Each stem cell contains one mutant gene copy and one 'wild-type' (normal) gene copy. The entire library is intended to mutate 13,000 genes in total. Of these 13000 mutant genes, 8000 mutations in mouse ES Cells are 'targeted': that is, the mutation which knocks out gene function is inserted precisely into the genome. The remaining 5000 mutations are derived from 'gene-traps'. These mutations are created by infecting ES Cells with an appropriately modified retrovirus. The retrovirus inserts itself 'randomly' into the mouse genome. When it inserts into a gene's intron, the introduced DNA will stop that gene copy being expressed, and the gene is considered to be knocked out (see gene trapping). [3] [4]

The mutations introduced in the ES Cells are conditional: this means that the initial mutation can be modified - by the application of particular DNA-altering enzymes (site specific recombinases) to make the knockout initially latent in the genome. The gene can be later knocked out (inactivated) at a specific time-point or tissue-type in mutant mice derived from the mutant ES Cells, by appropriate breeding to other transgenic mice. This conditionality is a key property of the entire resource, and it allows a more nuanced study of the effects of the gene-knockout.

Use of stem cells

The purpose of this library is to enable the mouse research community to rapidly create mutant mice, by

The broad aim of the project is to facilitate research into gene function in mice (and so by inference in humans) by removing the high technical entry barrier of making the mutant ES Cells.

The 8000 genes selected for mutation in mouse ES cells were drawn from the total set of protein coding mouse genes, based on a rough metric of how easy the genes were to mutate, with the technologies present at the time the project started. More 'target' genes were subsequently added to the list. All products from the program, as well links to the EUCOMM products for specific genes are available from the IKMC site. The distribution of EUCOMM products is being handled by a separate non-commercial organisation, the European Mouse Mutant Cell Repository.

Related Research Articles

Site-specific recombinase technologies are genome engineering tools that depend on recombinase enzymes to replace targeted sections of DNA.

Conditional gene knockout is a technique used to eliminate a specific gene in a certain tissue, such as the liver. This technique is useful to study the role of individual genes in living organisms. It differs from traditional gene knockout because it targets specific genes at specific times rather than being deleted from beginning of life. Using the conditional gene knockout technique eliminates many of the side effects from traditional gene knockout. In traditional gene knockout, embryonic death from a gene mutation can occur, and this prevents scientists from studying the gene in adults. Some tissues cannot be studied properly in isolation, so the gene must be inactive in a certain tissue while remaining active in others. With this technology, scientists are able to knockout genes at a specific stage in development and study how the knockout of a gene in one tissue affects the same gene in other tissues.

GSK3A

Glycogen synthase kinase-3 alpha is an enzyme that in humans is encoded by the GSK3A gene.

ARID2

AT-rich interactive domain-containing protein 2 (ARID2) is a protein that in humans is encoded by the ARID2 gene.

SUPT7L

STAGA complex 65 subunit gamma is a protein that in humans is encoded by the SUPT7L gene.

In molecular cloning and biology, a gene knock-in refers to a genetic engineering method that involves the one-for-one substitution of DNA sequence information in a genetic locus or the insertion of sequence information not found within the locus. Typically, this is done in mice since the technology for this process is more refined and there is a high degree of shared sequence complexity between mice and humans. The difference between knock-in technology and traditional transgenic techniques is that a knock-in involves a gene inserted into a specific locus, and is thus a "targeted" insertion. It is the opposite of gene knockout.

The ciliary neurotrophic factor receptor, also known as CNTFR, binds the ciliary neurotrophic factor. This receptor and its cognate ligand support the survival of neurons. This receptor is most closely related to the interleukin-6 receptor. This receptor possesses an unusual attachment to the cell membrane through a glycophosphatidylinositol linkage.

Knockout rat

A knockout rat is a genetically engineered rat with a single gene turned off through a targeted mutation used for academic and pharmaceutical research. Knockout rats can mimic human diseases and are important tools for studying gene function and for drug discovery and development. The production of knockout rats was not economically or technically feasible until 2008.

The International Knockout Mouse Consortium (IKMC) is a scientific endeavour to produce a collection of mouse embryonic stem cell lines that together lack every gene in the genome, and then to distribute the cells to scientific researchers to create knockout mice to study. Many of the targeted alleles are designed so that they can generate both complete and conditional gene knockout mice. The IKMC was initiated on March 15, 2007 at a meeting in Brussels. By 2011, Nature reported that approximately 17,000 different genes have already been disabled by the consortium, "leaving only around 3,000 more to go".

SLX4 protein involved in DNA repair

SLX4 is a protein involved in DNA repair, where it has important roles in the final steps of homologous recombination. Mutations in the gene are associated with the disease Fanconi anemia.

OTUD6B

OTU domain containing 6B is a protein that in humans is encoded by the OTUD6B gene.

<i>BBX</i> (gene)

HMG box transcription factor BBX also known as bobby sox homolog or HMG box-containing protein 2 is a protein that in humans is encoded by the BBX gene.

A knockout mouse, or knock-out mouse, is a genetically modified mouse in which researchers have inactivated, or "knocked out", an existing gene by replacing it or disrupting it with an artificial piece of DNA. They are important animal models for studying the role of genes which have been sequenced but whose functions have not been determined. By causing a specific gene to be inactive in the mouse, and observing any differences from normal behaviour or physiology, researchers can infer its probable function.

RNASEH2B

Ribonuclease H2, subunit B is a protein that in humans is encoded by the RNASEH2B gene. RNase H2 is composed of a single catalytic subunit (A) and two non-catalytic subunits, and degrades the RNA of RNA:DNA hybrids. The non-catalytic B subunit of RNase H2 is thought to play a role in DNA replication.

Malignant Brain Tumor domain containing 1 is a protein that in humans is encoded by the MBTD1 gene. The gene is also known as SA49P01.

Protein arginine methyltransferase 7 is a protein that in humans is encoded by the PRMT7 gene. Arginine methylation is an apparently irreversible protein modification catalyzed by arginine methyltransferases, such as PMT7, using S-adenosylmethionine (AdoMet) as the methyl donor. Arginine methylation is implicated in signal transduction, RNA transport, and RNA splicing.

RNA (guanine-9-) methyltransferase domain containing 2 is a protein that in humans is encoded by the RG9MTD2 gene. The gene is also known as TRM10.

Family with sequence similarity 104, member A is a protein that in humans is encoded by the FAM104A gene. The orthologous gene in mice is also known as D11Wsu99e.

Chromosome 5 open reading frame 45 is a protein that in humans is encoded by the C5orf45 gene. The orthologue in mice is 3010026O09Rik.

Mutagenesis (molecular biology technique)

In molecular biology, mutagenesis is an important laboratory technique whereby DNA mutations are deliberately engineered to produce libraries of mutant genes, proteins, strains of bacteria, or other genetically modified organisms. The various constituents of a gene, as well as its regulatory elements and its gene products, may be mutated so that the functioning of a genetic locus, process, or product can be examined in detail. The mutation may produce mutant proteins with interesting properties or enhanced or novel functions that may be of commercial use. Mutant strains may also be produced that have practical application or allow the molecular basis of a particular cell function to be investigated.

References

  1. ↑ International Mouse Knockout Consortium (2007). "A mouse for all reasons". Cell. 128 (1): 9–13. doi: 10.1016/j.cell.2006.12.018 . PMID   17218247.
  2. 1 2 van der Weyden L, White JK, Adams DJ, Logan DW (2011). "The mouse genetics toolkit: revealing function and mechanism". Genome Biol. 12 (6): 224. doi:10.1186/gb-2011-12-6-224. PMC   3218837 . PMID   21722353.
  3. ↑ Friedel RH, Seisenberger C, Kaloff C, Wurst W (2007). "EUCOMM--the European conditional mouse mutagenesis program". Brief Funct Genomic Proteomic. 6 (3): 180–5. doi: 10.1093/bfgp/elm022 . PMID   17967808.
  4. ↑ Bockamp E, Sprengel R, Eshkind L, Lehmann T, Braun JM, Emmrich F, Hengstler JG (2008). "Conditional transgenic mouse models: from the basics to genome-wide sets of knockouts and current studies of tissue regeneration". Regen Med. 3 (2): 217–35. doi:10.2217/17460751.3.2.217. PMID   18307405.
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