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| Rapid Construction of GPR126 Conditional Gene-targeting Vector |
| YE Xiang-li1,2, LI Da-li2 |
1. College of Medicine, Hunan Normal University, Changsha 410013, China;
2. Institute of Biomedical Sciences, East China Normal University, Shanghai 200241, China |
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Abstract Generation of specific gene knockout mouse model is a reliable technique for studying the role of genes in mammalian model organism. One of the key steps to acquire a gene knockout mouse is to construct a targeting vector for homologous recombination in mouse embryonic stem cells. For some genes, the mutants will die in uteri owing to their critical roles in embryonic development, or the mutant mice may have different phenotypes according to the period of development and types of tissues. It is difficult to study these genes by the conventional knockout approaches. Thereby the conditional knockout strategy had been developed for its advantages to circumvent the embryonic lethality problem and to investigate gene function temporally and spatially. Using modified Red homologous recombineering system, the two LoxP sites were inserted into the target position accurately and the GPR126 conditional gene-targeting vector was rapidly constructed. The successful generation of GPR126 conditional knockout construct will be helpful for the subsequent production of knockout mouse model and exploration of the function of GPR126 in mice.
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Received: 04 July 2012
Published: 25 April 2013
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[1] Capecchi M R. Targeted gene replacement. Sci Am, 1994, 270: 52-59.
[2] Li J, Baker M D. Mechanisms involved in targeted gene replacement in mammalian cells. Genetics, 2000, 156: 809-821.
[3] Yin H F, Wang Q J, Li N. The research progress in the disease model of knock-out mouse. Yi Chuan, 2002, 24: 463-469.
[4] Betz U A, Vosshenrich C A, Rajewsky K, et al. Bypass of lethality with mosaic mice generated by Cre-loxP-mediated recombination. Curr Biol, 1996, 6: 1307-1316.
[5] Pluck A. Conditional mutagenesis in mice: the Cre/loxP recombination system. Int J Exp Pathol, 1996, 77: 269-278.
[6] Sternberg N, Hamilton D. Bacteriophage P1 site-specific recombination. I. Recombination between loxP sites. J Mol Biol, 1981, 150: 467-486.
[7] Guo F, Gopaul D N, van Duyne G D. Structure of Cre recombinase complexed with DNA in a site-specific recombination synapse. Nature, 1997, 389: 40-46.
[8] Baud V, Chissoe S L, Viegas-Pequignot E, et al. EMR1, an unusual member in the family of hormone receptors with seven transmembrane segments. Genomics, 1995, 26: 334-344.
[9] Stehlik C, Kroismayr R, Dorfleutner A, et al. VIGR—a novel inducible adhesion family G-protein coupled receptor in endothelial cells. FEBS Lett, 2004, 569: 149-155.
[10] Monk K R, Naylor S G. A G protein-coupled receptor is essential for Schwann cells to initiate myelination. Science, 2009, 325: 1402-1405.
[11] Aguayo A J, Kasarjian J, Skamene E, et al. Myelination of mouse axons by Schwann cells transplanted from normal and abnormal human nerves. Nature, 1977, 268: 753-755.
[12] Sherman D L, Brophy P J. Mechanisms of axon ensheathment and myelin growth. Nat Rev Neurosci, 2005, 6: 683-690.
[13] Waller-Evans H, Promel S, Langenhan T, et al. The orphan adhesion-GPCR GPR126 is required for embryonic development in the mouse. PLoS One, 2010, 5: e14047.
[14] 姜丽, 叶湘漓, 李大力. 利用Red重组系统快速构建基因打靶载体. 中国生物工程杂志, 2011, 31(10): 88-94. Jiang L, Ye X L, Li D L. Construction of mouse gene-targeting vector through modified recombineering strategy. China Biotechnology, 2011, 31(10): 88-94.
[15] Bunting M, Bernstein K E, Greer J M, et al. Targeting genes for self-excision in the germ line. Genes Dev, 1999, 13: 1524-1528.
[16] Nakano M, Ishimura M, Chiba J, et al. DNA substrates influence the recombination efficiency mediated by FLP recombinase expressed in mammalian cells. Microbiol Immunol, 2001, 45: 657-665.
[17] Wu S, Ying G, Wu Q, et al. A protocol for constructing gene targeting vectors: generating knockout mice for the cadherin family and beyond. Nat Protoc, 2008, 3: 1056-1076.
[18] 王军平, 张友明. Red/ET重组在基因打靶载体快速构建中的应用. 遗传, 2005, 27(6): 953-958. Wang J P, Zhang Y M. The application of Red/ET recombination to high efficient gene-targeting vector construction.Yi Chuan, 2005, 27(6): 953-958.
[19] 周芳亮, 胡翔, 吴文韬, 等. GAS41基因在斑马鱼胚胎发育中的时空表达谱. 湖南师范大学自然科学学报, 2012, 35(1): 66-70. Zhou F L, Hu X, Wu W T, et al. Temporal and spatial expression profiling of GAS41 gene in zebrafish embryonic development.Journal of Natural Science of Hunan Normal University, 2012, 35(1): 66-70. |
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