TY - JOUR
T1 - Producing molecular biology reagents without purification
AU - Bhadra, Sanchita
AU - Nguyen, Vylan
AU - Torres, Jose Angel
AU - Kar, Shaunak
AU - Fadanka, Stéphane
AU - Gandini, Chiara
AU - Akligoh, Harry
AU - Paik, Inyup
AU - Maranhao, Andre C.
AU - Molloy, Jenny
AU - Ellington, Andrew D.
N1 - Publisher Copyright:
© 2021 Bhadra et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2021/6
Y1 - 2021/6
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/85106969415
UR - https://www.scopus.com/inward/citedby.url?scp=85106969415&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0252507
DO - 10.1371/journal.pone.0252507
M3 - Article
C2 - 34061896
AN - SCOPUS:85106969415
SN - 1932-6203
VL - 16
JO - PLoS ONE
JF - PLoS ONE
IS - 6 June
M1 - e0252507
ER -