MUSERGEN: MultiUSER equipment for GENe identification in biosciences and biotechnology

提高我们对生物学的理解在很大程度上是通过对基因序列（以及蛋白质）的自然或实验室操作，以及对感兴趣的生物体的影响的观察来完成的。这些影响包括对各种压力的耐受性，代谢感兴趣的化合物的能力，延长寿命的能力，保持健康的能力，等等。原则上，一个人可能会多次这样做，以产生大量的变体库。如果该生物体能够生产某种感兴趣的产品（如青霉素、生物燃料或其他任何东西），这也有可能显著提高宿主的生产力。在实践中，大多数方法只能去除（敲除）基因的活性，但很少有方法能够以分级的方式操纵它们，这往往是在定制或“工艺”行业中完成的。最近，人们发现了一种细菌对抗某些病毒的“免疫防御”系统，即CRISPR-Cas9，可以进行更精确的“基因编辑”，这种通用策略正在彻底改变生物学。然而，再一次，编辑往往是一次完成一个。就在最近（2019年10月1日），一种能够在全基因组范围内进行这种基因编辑的仪器问世了，可以同时有效地进行数千次这样的实验（并行）。它使用了一个带有自己专有系统的CRISPR-Cas9版本（但对商业用途完全免费）。通过用条形码标记每个已知的编辑，并评估特定条形码在感兴趣的条件下表达的频率，人们可以同时评估数百个基因的相对重要性，以及它们应该如何被修改。该仪器旨在使基于crispr的基因编辑“民主化”，它被称为Inscripta Onyx仪器。该提案的目的是获得资金购买这样一种仪器，将其安装在利物浦的合成生物学中心（基因组研究中心的基因工厂），并使其可供当地和更广泛的BBSRC社区使用。这个项目的结果将是大量的新知识，哪些基因-往往是意想不到的-参与各种感兴趣的生物过程。它将使我们能够以真正合理的方式控制和优化细胞酶和小分子的生产。

Improving our understanding of biology is largely accomplished by natural or laboratory manipulation of the sequences of genes (and hence proteins), and observations of the effects on the organisms of interest. Such effects can include tolerance to various stresses, the ability to metabolise compounds of interest, the ability to improve longevity, to remain healthy, and so on. In principle one might do this many times to produce vast libraries of variants. If the organism is one that makes a product of interest (e.g. penicillin, a biofuel, or indeed anything else), this also leads to the possibility of improving the host's productivity dramatically. In practice, most methods are able only to remove (knock out) the activity of genes, but fewer can be used to manipulate them in a graded manner, and this tends to be done in a bespoke or 'craft' industry kind of way. More recently, it was discovered that a system of 'immune defence' for bacteria against certain viruses, known as CRISPR-Cas9, allows a much more precise 'gene editing', and this general strategy is revolutionising biology. Again, however, the edits tend to be done one at a time.Very recently (October 1st, 2019), an instrument became available that is able to perform such gene editing on a genome-wide scale, allowing thousands of such experiments effectively to be done at once (in parallel). It uses a version of CRISPR-Cas9 with its own proprietary system (but one that is entirely free for commercial use). By tagging each known edit with a barcode, and assessing the frequency with which particular barcodes are expressed under a condition of interest, one can assess the relative importance of hundreds of genes simultaneously, and how they should be modified. This instrument is designed to 'democratise' CRISPR-based gene editing, and it is known as the Inscripta Onyx instrument. The purpose of this proposal is to obtain funds to purchase such an instrument, to set it up in Liverpool's synthetic biology centre (the Gene Mill in the Centre for Genomic Research), and to make it available to the local and wider BBSRC community.The result of this program will be huge amounts of new knowledge of which genes - which will often be unexpected ones - are involved in various bioprocesses of interest. It will allow us to control and optimize cellular enzyme and small molecule production in a truly rational manner.