This invention relates to in vitro production of nucleic acids, particularly RNAs and specifically messenger RNAs (mRNA).

The present disclosure provides recombinant microorganisms capable of sugar co-utilization, the recombinant microorganism comprising a genetically altered microorganism (e.g. S. cerevisiae) having a lack of, or reduced expression of, or expression of truncated or mutated forms of at least one polypeptide selected from Hxk1, Hxk2, and Glk1. Reduction of the expression or biological activity of Hxk1, Hxk2 and Glk1 leads to reduction in the glucose consumption pathway, allowing the microorganism to co-utilize multiple sugars (e.g. glucose, xylose, galactose) at an improved rate.

The present invention provides compositions, kits, and methods for synthesizing polyadenylic acid using polynucleotide phosphorylase, adenosine diphosphate, a buffering agent, and a divalent metal cation. In certain embodiments, the adenosine diphosphate is present at a concentration between about 5.0 mM and about 100 mM, and the buffering agent has a pH around 8.0. In some embodiments, the prepared polyadenylic acid compositions are free of detectable nucleic acid.

Methods and compositions for genetically modifying a cell are provided.