Synopsis of Project

In this project we will be using CRISPR-Cas9 nickase and homologous recombination to duplicate and delete native genes to make Synechococcus elogus 7002 pump salt from the media into the intracellular space. Synechococcus can grow in a large range of salinic conditions, from freshwater to seawater. The genes that give  the ability for Synechococcus to achieve this feat are the genes we are targeting. We are at Phase 1.

Engineering challenges

1. Salt kills metabolism
2. Use of endogenous genes exclusively
3. Filtering bacteria post treatment
4. Disposal of waste product

Phase 1

Directed mutagenesis of wild type strain to produce a more salt tolerant strain.
Comparison CRISPR-Cas9 transformation efficiency to conventional homologous recombination.

Phase 2

Deletion of genes that transport salt out and allow water in.
Flip the direction of salt exporters so they become importers.
Duplicate membrane synthesis and salt tolerances genes.

Phase 3

Designing system scale up and optimization.

Lab Notebook

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