Assistant Prof. @uoknightcampus.bsky.social. Cofounder @synplexity.bsky.social. Gene synthesis, synbio, protein engineering, nanopores, multiplex assays. Opinions my own. www.plesalab.org
Calin Plesa
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We’re excited to announce the next Fluorescent Proteins & Biological Sensors conference! Abstract submissions are now open—hope to see you there.
www.janelia.org/you-janelia/...
Just released! Fluorescent Protein Libraries from the Plesa Lab. Benchmarking protein expression, developing imaging tools, fluorescent protein engineering, and more.
www.addgene.org/pooled-libra...
Traditional DropSynth uses PCA, which becomes increasingly prone to cross-hybridization artifacts, partial assemblies, and amplification bias especially as fragment count increases. 2/n
We kept everything that makes DropSynth scalable: barcoded bead capture, emulsion compartmentalization, and pooled processing, but replaced PCA with Type IIS Golden Gate Assembly inside droplets. 3/n
Many GGA library assembly methods are limited by the number of orthogonal junctions that can coexist in a shared reaction. DropSynth-Gold performs each assembly in its own droplet. Library scale is determined primarily by barcode diversity, not junction diversity. 5/n
We're still pushing DS on length and scale with more in the pipeline. In the meantime, check the preprint for lots more details on DS-Gold: www.biorxiv.org/content/10.6...
We benchmarked 6 libraries spanning 5-12 fragments per gene corresponding to lengths of ~1 kb to ~3 kb. GGA had comparable coverage and fidelity with a slight hit on uniformity. Fidelity (1 in ~1200 bp) remained stable across all lengths. 4/n
We just preprinted DropSynth-Gold where we replaced PCA with Golden Gate Assembly (GGA) inside emulsion droplets, extending multiplexed gene synthesis to ~3 kb constructs assembled from up to 12 fragments while preserving the scalability of DropSynth. 1/n
