Producing molecular biology reagents without purification.

We recently developed 'cellular' reagents-lyophilized bacteria overexpressing proteins of interest-that can replace commercial pure enzymes in typical diagnostic and molecular biology reactions. To make cellular reagent technology widely accessible and amenable to local production with minimal instrumentation, we now report a significantly simplified method for preparing cellular reagents that requires only a common bacterial incubator to grow and subsequently dry enzyme-expressing bacteria at 37°C with the aid of inexpensive chemical desiccants. We demonstrate application of such dried cellular reagents in common molecular and synthetic biology processes, such as PCR, qPCR, reverse transcription, isothermal amplification, and Golden Gate DNA assembly, in building easy-to-use testing kits, and in rapid reagent production for meeting extraordinary diagnostic demands such as those being faced in the ongoing SARS-CoV-2 pandemic. Furthermore, we demonstrate feasibility of local production by successfully implementing this minimized procedure and preparing cellular reagents in several countries, including the United Kingdom, Cameroon, and Ghana. Our results demonstrate possibilities for readily scalable local and distributed reagent production, and further instantiate the opportunities available via synthetic biology in general.

Déclaration de conflit d'intérêts

HA is affiliated with Kumasi Hive, a Ghanaian not-for-profit social enterprise. SF is Executive Director of Mboalab, a Cameroonian registered company. JM is an unpaid Executive Director of Beneficial Bio Ltd, a UK not-for-profit company limited by guarantee and a volunteer board member of the Gathering for Open Science Hardware Inc, a US 501c3 nonprofit. ADE and SB are named inventors on pending US patent application S20200399679A1 covering cellular reagents, assigned to the University of Texas System. These competing interests do not alter our adherence to PLOS ONE policies on sharing data and materials.