GreenGate 2.0

Very happy to introduce GreenGate 2.0 (GG2.0)  a backwards-compatible addon for the assembly of complex transcriptional units and their stacking. 

Modular molecular cloning is in genetic engineering, enabling precise design of synthetic transcriptional units (TUs) & genome manipulation. GreenGate has been very popular, but it had limitation.

TU could be assembled from a max of 6 modules. Generating module was done by classical restriction/ligation. Creating multi-TU T-DNA was cumbersome and inefficient. GG2.0 tackles these issues: GG2.0 allows up to 12 Level 0 modules in a single Level 1 TU, opening doors for more complex applications like multiplex genome editing.

GG2.0 allows the easy generation of Level 0 module thanks to a Universal Entry Generator plasmid (pUEG). One-pot, one-step reactions now feasible for entry clones and domesticate internal sites!

GreenBraid, a new feature in GG2.0, enables iterative stacking of transcriptional units for multigene assemblies. Say goodbye to inefficient sequential Level 2 assembly! Good news! GG2.0 remains compatible with most existing GreenGate modules, making adoption easier for researchers. 

And that's not all! GG2.0 introduces Level 0 modules for CRISPR/Cas9 guide RNA expression & pre-assembled Level 1 vectors for dexamethasone-inducible gene expression & nuclear + plasma membrane fluorescent markers.

GG 2.0 will soon be available on AddGene!  

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.02900…