Self-assembly of a multimeric genomic hydrogel via multi-primed chain reaction of dual single-stranded circular plasmids for cell-free protein production

Hyangsu Nam; Taehyeon Kim; Sunghyun Moon; Yoonbin Ji; Jong Bum Lee

doi:10.1016/j.isci.2023.107089

Self-assembly of a multimeric genomic hydrogel via multi-primed chain reaction of dual single-stranded circular plasmids for cell-free protein production

iScience. 2023 Jun 10;26(7):107089. doi: 10.1016/j.isci.2023.107089. eCollection 2023 Jul 21.

Authors

Hyangsu Nam¹, Taehyeon Kim¹, Sunghyun Moon¹, Yoonbin Ji¹, Jong Bum Lee¹

Affiliation

¹ Department of Chemical Engineering, University of Seoul, 163 Seoulsiripdaero, Dongdaemun-gu, Seoul 02504, Republic of Korea.

Abstract

Recent technical advances in cell-free protein synthesis (CFPS) offer several advantages over cell-based expression systems, including the application of cellular machinery, such as transcription and translation, in the test tube. Inspired by the advantages of CFPS, we have fabricated a multimeric genomic DNA hydrogel (mGD-gel) via rolling circle chain amplification (RCCA) using dual single-stranded circular plasmids with multiple primers. The mGD-gel exhibited significantly enhanced protein yield. In addition, mGD-gel can be reused at least five times, and the shape of the mGD-gel can be easily manipulated without losing the feasibility of protein expression. The mGD-gel platform based on the self-assembly of multimeric genomic DNA strands (mGD strands) has the potential to be used in CFPS systems for a variety of biotechnological applications.

Keywords: Biotechnology; Molecular self-assembly; Scaffolds in supramolecular chemistry.