Abstract: A method for cell-free protein synthesis, the method comprising synthesizing RNA or a protein of interest in a reaction mixture comprising: a template DNA encoding the RNA or protein of interest; and a biological extract of a protease-deficient bacterial cell that has been genetically modified to express a heterologous RNA polymerase, the extract further comprising components necessary for transcription and translation of the protein. Also provided are a method for producing a reaction mixture for cell-free protein synthesis and kits for executing these methods.