Team:Tianjin/pz

Design

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Design

The project we want to accomplish is a chromosomal stabilization element group centered on centromeres, which transfer large DNA fragments by transferring chromosomes. The difference between the source host and the destination host of the chromosome may be very large[1], so we need to enhance the stability of the chromosome.

Chromosomal stabilization element group

In the application of our chromosome-based gene transfer technology to the actual process, it is highly probable that the objective chromosome and the chromosome of Saccharomyces cerevisiae are far apart. At this time, in order to ensure the stable existence and stable inheritance of our large fragments after introduction into host cells, we attempt to introduce chromosomal stabilization element group on chromosomes with large fragments.

1. Centromere introduction

So we adjusted the experiment: using the centromere of the E chromosome of Y. lipolytica to replace the centromere of synthetic chromosome V of S. cerevisiae, and then the strain we obtained grows well.Click here to learn more.

In addition, we will transfer the synthetic chromosome V with lycopene pathway on this strain to Y. lipolytica. Because Y. lipolytica's naturally occurring liposomes are suitable for enrichment of hydrophobic products, and their ability to utilize cheap raw materials can help us build high-yield Y. lipolytica strains faster, and then optimize yields with the help of a genomic rearrangement system which are unique to synthetic chromosomes.

2. Chromosome transfer

On the other hand, we replaced the centromere of S.cerevisiae with the centromere of Y. lipolytica, so that when we transfer this chromosome into Y. lipolytica, it not only has the characteristics of being able to use cheap raw materials, but also has all genes that needed in the lycopene peoduction. Appart from this, it can store these lycopene with its abundant liposomes which can achieve a higher production than S. cerevisiae. After that, we will perform SCRaMbLE on the synthetic chromosome V in Y. lipolytica to achieve high-throughput gene replication, deletion, inversion, and translocation, thereby further increasing the yield.^[4]

Figure 8. Schematic diagram of SCRaMbLE

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Design

The project we want to accomplish is a chromosomal stabilization element group centered on centromeres, which transfer large DNA fragments by transferring chromosomes. The difference between the source host and the destination host of the chromosome may be very large[1], so we need to enhance the stability of the chromosome.

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Reference

[1] Dujon Bernard. Yeast evolutionary genomics.[J]. Nature Reviews. Genetics,2010,11(7).

[2] Gautam Chatterjee, Sundar Ram Sankaranarayanan, Krishnendu Guin,etc.Repeat-Associated Fission Yeast-Like Regional Centromeres in the Ascomycetous Budding Yeast Candida tropicalis[J].PLOS Genetics,2016:1-28.

[3] Amor DJ, Kalitsis P, Sumer H, Choo KH (2004) .Building the centromere:from foundation proteins to 3D organization.Trends Cell Biol14, 359-368.

[4] "Perfect" designer chromosome V and behavior of a ring derivative. Science 355, eaaf4704 (2017).

[5] Gaudelli N M , Komor A C , Rees H A , et al. Publisher Correction: Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage[J]. Nature, 2018, 559(7714):E8.