M13 bacteriophage is a well-characterized platfor
m for peptide display. The utility of the M13 display platfor
m is derived fro
m the ability to encode phage protein fusions with display peptides at the geno
mic level. However, the geno
me of the phage is co
mplicated by overlaps of key genetic ele
ments. These overlaps directly couple the coding sequence of one gene to the coding or regulatory sequence of another,
making it difficult to alter one gene without disrupting the other. Specifically, overlap of the end of gene VII and the beginning of gene IX has prevented the functional geno
mic
modification of the N-ter
minus of p9. By redesigning the M13 geno
me to physically separate these overlapping genetic ele
ments, a process known as 鈥渞efactoring,鈥?we enabled independent
manipulation of gene VII and gene IX and the construction of the first N-ter
minal geno
mic
modification of p9 for peptide display. We de
monstrate the utility of this refactored geno
me by developing an M13 bacteriophage-based platfor
m for targeted i
maging of and drug delivery to prostate cancer cells
in vitro. This successful use of refactoring principles to re-engineer a natural biological syste
m strengthens the suggestion that natural geno
mes can be rationally designed for a nu
mber of applications.
Keywords:
me+refactoring&qsSearchArea=searchText">genome refactoring; drug delivery; M13 bacteriophage; synthetic biology