New recombinases for genome engineering

用于基因组工程的新重组酶

• 批准号: BB/H005277/1
• 负责人: William Brown
• 金额: $ 43.09万
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2010
• 资助国家: 英国
• 起止时间: 2010 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FH005277%2F1
• 关键词: New; recombinases; genome; engineering

Site-specific recombinases are a class of proteins that splice different DNA sequences together in a specific and predictable way. They are widely used in biotechnology to manipulate the genomes of plants and animals. A particularly useful application of these proteins is to insert or delete genes or segments of DNA in mammalian chromosomes to study their function. Ideally inserted DNA should be prevented from excising (and excised DNA must be prevented from re-inserting) and this can be done with one group of site-specific recombinases called the serine integrases. One of us, Prof Smith, discovered that the integrase from a bacterial virus, phiC31 can promote an irreversible insertion or deletion without the participation of any other protein. This protein has, as a consequence, become very widely used in nearly all branches of experimental biology. phiC31 integrase is not perfect however and in fact suffers from major problems. Sometimes the reaction promoted by phiC31 integrase is incomplete and sometimes it does not happen at all. Many other serine recombinases have been discovered in the course of genome projects and some may be much better than the original phiC31 integrase for use in biotechnology. Surprisingly these new proteins have never been studied systematically in order to determine whether any of them are in fact better than phiC31 integrase. We would like to do this.

位点特异性重组酶是一类以特定和可预测的方式将不同的DNA序列拼接在一起的蛋白质。它们被广泛用于生物技术，以操纵植物和动物的基因组。这些蛋白质的一个特别有用的应用是插入或删除哺乳动物染色体中的基因或DNA片段，以研究它们的功能。理想情况下，插入的DNA应该防止切除(切除的DNA必须防止重新插入)，这可以通过一组称为丝氨酸整合酶的位点特异性重组酶来完成。我们中的一位，史密斯教授，发现了来自细菌病毒phiC31的整合酶可以在没有任何其他蛋白质参与的情况下促进不可逆转的插入或缺失。因此，这种蛋白质在实验生物学的几乎所有分支中都得到了非常广泛的应用。然而，phiC31整合酶并不是完美无缺的，事实上存在着重大问题。有时phiC31整合酶促进的反应是不完全的，有时根本不发生。在基因组计划的过程中已经发现了许多其他丝氨酸重组酶，其中一些可能比最初用于生物技术的phiC31整合酶要好得多。令人惊讶的是，这些新的蛋白质从来没有被系统地研究过，以确定它们中是否有任何一个实际上比phiC31整合酶更好。我们想做这件事。

项目成果

Accuracy and efficiency define Bxb1 integrase as the best of fifteen candidate serine recombinases for the integration of DNA into the human genome.

• DOI: 10.1186/1472-6750-13-87
• 发表时间: 2013-10-20
• 期刊: BMC biotechnology
• 影响因子: 3.5
• 作者: Xu Z;Thomas L;Davies B;Chalmers R;Smith M;Brown W
• 通讯作者: Brown W

Comparison and optimization of ten phage encoded serine integrases for genome engineering in Saccharomyces cerevisiae.

• DOI: 10.1186/s12896-016-0241-5
• 发表时间: 2016-02-09
• 期刊: BMC biotechnology
• 影响因子: 3.5
• 作者: Xu Z;Brown WR
• 通讯作者: Brown WR

Rapid metabolic pathway assembly and modification using serine integrase site-specific recombination.

• DOI: 10.1093/nar/gkt1101
• 发表时间: 2014-02
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Colloms SD;Merrick CA;Olorunniji FJ;Stark WM;Smith MC;Osbourn A;Keasling JD;Rosser SJ
• 通讯作者: Rosser SJ