Cell-free production of ribonucleic acid

A cell-free method of biosynthesizing ribonucleic acid (RNA), the method comprising:

• (a) incubating a cell lysate mixture that comprises (i) cellular RNA and (ii) an enzyme that depolymerizes RNA, and (iii) a thermostable kinase and/or a thermostable RNA polymerase;
• and producing, a cell lysate mixture that comprises nucleoside monophosphates;
• (b) heating the cell lysate mixture produced in step (a) to a temperature that inactivates or partially inactivates the enzymes that depolymerize RNA and enzymes that degrade nucleotides and/or nucleic acids without completely inactivating the thermostable kinase activities and/or thermostable RNA polymerase activities, and producing a cell lysate mixture that comprises nucleoside monophosphates, and the thermostable kinase and/or the RNA polymerase;
• and
• (c) incubating the cell lysate mixture produced in (b) in the presence of an energy source and a deoxyribonucleic acid (DNA) template encoding a RNA of interest, producing nucleoside triphosphates, and producing a cell lysate mixture that comprises the RNA of interest.