From Electronic Sequence to Purified Protein Using Automated Gene Synthesis and In Vitro Transcription/Translation

• DOI: 10.1021/acssynbio.0c00060
• Publication Date: 2020-06-05T21:45:16Z
• Authors: Claudia Chiocchini; Krishna Vattem; Michael Liss; Lisa Ludewig; Tobias Reusch; Ischwaku Rastogi; Brian Webb; Axel Trefzer
• Keywords: Synthetic Biology; Cell-free protein synthesis; Cloning (programming); Transcription

De novo gene synthesis is the state-of-the-art method used to obtain genetic material adapted requirements of host organism and a cornerstone for modern synthetic biology. Yet, little progress has been made regarding downstream processes protein production from material. The recombinant proteins traditionally requires extensive preparatory work including amplification, cloning, sequencing, transformation or transfection expression host, cultivation living cells, purification overexpressed protein. In this we describe fast automated workflow cell-free starting an electronic sequence accession number. PRESTO (protein oligonucleotides) seamlessly combines tailored in silico optimization with assembly short oligonucleotides into linear DNA cassettes, mammalian vitro transcription/translation, thereof. Integrated on small liquid handling system it provides hands-free high throughput source functional within 1 day.