For example, the temperature-sensitive lethal genetic screens performed in the 1960s and 70s in microbes were essential initial steps in providing a foundation for our current understanding of how DNA replication, transcription, and translation take place within the cell. Genetics has been, and will continue to be, an essential tool for providing insight into molecular and biological function in all forms of life. The ability to perform precision genome engineering in medically and industrially important lactic acid bacteria will allow for the genetic improvement of strains without compromising safety. The emergence of CRISPR-Cas technology in genome engineering and its potential application to enhancing SSDR in lactic acid bacteria is discussed. We also provide a blueprint for how to proceed if one is attempting to establish SSDR technology in a lactic acid bacterium. coli into Lactobacillus reuteri and Lactococcus lactis. In this review we discuss the application of SSDR technology in lactic acid bacteria, with an emphasis on key factors that were critical to move this technology from E. Single-stranded DNA recombineering (SSDR) allows for the generation of subtle mutations without the need for selection and without leaving behind any foreign DNA. One of these technologies, recombination mediated genetic engineering (recombineering) allows for precision DNA engineering of chromosomes and plasmids in Escherichia coli. Innovative new genome engineering technologies for manipulating chromosomes have appeared in the last decade.
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