Broad host range genetic tools for engineering microalgae
Disclosed herein are a suite of genetic tools suitable for engineering both the nucleus and chloroplast in diverse microalgae.
1. A method for transforming a Picochlorum renovo algal chloroplast, the method comprising:
introducing an exogenous selection marker into the Picochlorum renovo algal chloroplast; and
bombarding a culture of the Picochlorum renovo algal chloroplast with a loaded particle.
2. A method for transforming a Picochlorum renovo algal nucleus, the method comprising:
synthesizing a nuclear integration cassette;
integrating the nuclear integration cassette into a plasmid backbone; and
codon optimizing a gene encoding a selection marker and a gene encoding a peptide into a genome of the Picochlorum renovo algal nucleus;
bombarding a culture of the Picochlorum renovo algal nucleus with a loaded particle.
3. The method of claim 2 , wherein the selection marker encodes for resistance to erythromycin.
4. The method of claim 2 , wherein the plasmid backbone is pUC19.
5. The method of claim 2 , wherein the peptide is a 2A peptide.
6. The method of claim 2 , wherein the loaded particle comprises:
a gold sphere nanoparticle,
plasmid DNA precipitated onto the gold sphere nanoparticle, and
spermidine.
7. The method of claim 1 , wherein the selection marker encodes for resistance to erythromycin.
8. The method of claim 1 , wherein the loaded particle comprises:
a gold sphere nanoparticle,
plasmid DNA precipitated onto the gold sphere nanoparticle, and
spermidine.