Cell-free production of ribonucleic acid

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

• (a) incubating a cell lysate mixture that comprises cellular RNA, an enzyme that depolymerizes RNA, a thermostable kinase, and 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 kinases and thermostable RNA polymerases, and producing a cell lysate mixture that comprises nucleoside monophosphates, a thermostable kinase, and a 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.