Methods and compositions for performing continuous directed evolution

A method of performing continuous directed evolution of a polynucleic acid sequence, wherein said polynucleic acid sequence comprises the sequence of a gene of interest, said method comprising:

• (a) contacting a plurality of engineered, non-naturally occurring mammalian cells with a plurality of engineered, non-naturally occurring adenoviruses capable of infecting the mammalian cells, thereby producing a population of infected cells, wherein:
• (i) each engineered, non-naturally occurring adenovirus comprises a modified genome, wherein the modified genome comprises:
• an integration of a polynucleic acid sequence comprising the sequence of a transcribable gene of interest, a deletion of the sequence encoding for the adenoviral DNA polymerase, and a deletion of a sequence encoding for a protein necessary for the production of infectious adenovirus particles wherein the protein necessary for the production of infectious adenovirus particles is not the adenoviral DNA polymerase;
• and
• (ii) each engineered, non-naturally occurring mammalian cell comprises a modified genome, wherein the modified genome comprises:
• an integration of a polynucleic acid sequence comprising the sequence of an error-prone adenoviral DNA polymerase, and an integration of a polynucleic acid sequence comprising the sequence encoding for the protein necessary for the production of infectious adenovirus particles in (a)(i) under the control of a functional couple, optionally wherein the functional couple is a transcriptionally-coupled promoter, and
• (b) generating infectious adenovirus particles with the infected cells, wherein the generation of the infectious adenovirus particles is increased by the evolution of the polynucleic acid sequence comprising the sequence of the transcribable gene of interest wherein said evolution is driven by the error-prone adenoviral DNA polymerase.