musebio

Genome scale engineering

Kevin Ness, CEO, Muse Bio

bd@musebio.com

https://www.youtube.com/watch?v=qeb0KC9bs98&t=0s&list=PLHpV_30XFQ8RN0v_PIiPKnf8c_QHVztFM&index=56

Single cell writing

Sold a digital PCR company to Agilent(?). Started 10x genomics. Those were focused on better tools to read the genome. Now the focus is better tools to write the genome. I can't do this alone, I am a tool builder not a tool user. We are inserting barcodes so that you can track each cell. We started in 2015 from U of Colorado. 6M series A. 23M series B from lead investors in intel and illumina. We just announced founder/chairman of illumina joining our board. We want to write the genome. Expected commercial launch end of 2018.

Single cell writing with trackable multiplex editing

Some of this was published in Nature Biotech brand name will be ForgeCraft biochemistry. "Create" (?).

96 well plate is limited. You are limited to the number of wells. If you want to do an edit with a guide RNA and so on, and you need more wells. Whether you need a library of guide RNA, or a library of homology arms, to try to increase the number of edits per experiment, you are limited by the plate. Don't think of using the well as the partition. Let's use the cell as the partition. Using the cell as the partition, you can put 100k well plate in a normal 96 well plate. How do you get the reagents colocalized always going into the right cell?

So they are covalently attached-- gRNA and homology arm are combined. Also, you can trakc it, so there's a barcode attached using biochemistry.

We rationally designed every variant for the FolA enzyme. We screened activity to find viable variants. 100x reduction in cost, 200x increase in activity, >3140 mutants, novel mutations identified. This is a well studied protein but we found novel variants because we're doing full saturation of the enzyme, literally every combination of the amino acid sequence. So let's jump from one gene and go to 19 genes.

Here you're now studying a pathway. We wanted to increase the lysine production. We got 100x increase in variation space with 10x reduction in time. There was a 1000x increase in growth. 16300 variants, 19 genes. We are doing mass spec verification.

We also did a network with 177 genes. We made 75,093 variants. 64x increase in antibiotic resistance was discovered.

ForgeCrafter (software), Forge (instrument), and Forge kits (reagents). Fully automated plasmid and single cell gene writing with barcodes.