MUSERGEN: MultiUSER equipment for GENe identification in biosciences and biotechnology

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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.