Multiplexed genome engineering and genotyping methods applications for synthetic biology and metabolic engineering

Methods Enzymol. 2011;498:409-26. doi: 10.1016/B978-0-12-385120-8.00018-8.

Abstract

Engineering at the scale of whole genomes requires fundamentally new molecular biology tools. Recent advances in recombineering using synthetic oligonucleotides enable the rapid generation of mutants at high efficiency and specificity and can be implemented at the genome scale. With these techniques, libraries of mutants can be generated, from which individuals with functionally useful phenotypes can be isolated. Furthermore, populations of cells can be evolved in situ by directed evolution using complex pools of oligonucleotides. Here, we discuss ways to utilize these multiplexed genome engineering methods, with special emphasis on experimental design and implementation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • Chromosomes / genetics
  • Genetic Engineering / methods*
  • Genome*
  • Genotype*
  • Molecular Biology / methods
  • Molecular Sequence Data
  • Oligonucleotides / genetics
  • Polymerase Chain Reaction / methods
  • Recombination, Genetic
  • Synthetic Biology / methods*

Substances

  • Oligonucleotides