RESUMO
The Type II CRISPR-Cas9 system is a simple, efficient, and versatile tool for targeted genome editing in a wide range of organisms and cell types. It continues to gain more scientific interest and has established itself as an extremely powerful technology within our synthetic biology toolkit. It works upon a targeted site and generates a double strand breaks that become repaired by either the NHEJ or the HDR pathway, modifying or permanently replacing the genomic target sequences of interest. These can include viral targets, single-mutation genetic diseases, and multiple-site corrections for wide scale disease states, offering the potential to manage and cure some of mankind's most persistent biomedical menaces. Here, we present the developing progress and future potential of CRISPR-Cas9 in biological and biomedical investigations, toward numerous therapeutic, biomedical, and biotechnological applications, as well as some of the challenges within. J. Cell. Biochem. 119: 81-94, 2018. © 2017 Wiley Periodicals, Inc.
Assuntos
Sistemas CRISPR-Cas , Edição de Genes , Animais , Infecções Bacterianas/terapia , Genética Microbiana/história , Genômica , História do Século XX , História do Século XXI , Humanos , Modelos Animais , Neoplasias/terapia , Terapêutica , Viroses/terapiaAssuntos
Genética Microbiana/métodos , Biologia Molecular/métodos , Micologia/métodos , Saccharomyces/genética , Saccharomyces/fisiologia , Genética Microbiana/história , Genética Microbiana/tendências , História do Século XX , História do Século XXI , Humanos , Biologia Molecular/história , Biologia Molecular/tendências , Micologia/história , Micologia/tendênciasRESUMO
Budding yeast strains used in the laboratory have had a checkered past. Historically, the choice of strain for any particular experiment depended on the suitability of the strain for the topic of study (e.g., cell cycle vs. meiosis). Many laboratory strains had poor fermentation properties and were not representative of the robust strains used for domestic purposes. Most strains were related to each other, but investigators usually had only vague notions about the extent of their relationships. Isogenicity was difficult to confirm before the advent of molecular genetic techniques. However, their ease of growth and manipulation in laboratory conditions made them "the model" model organism, and they still provided a great deal of fundamental knowledge. Indeed, more than one Nobel Prize has been won using them. Most of these strains continue to be powerful tools, and isogenic derivatives of many of them-including entire collections of deletions, overexpression constructs, and tagged gene products-are now available. Furthermore, many of these strains are now sequenced, providing intimate knowledge of their relationships. Recent collections, new isolates, and the creation of genetically tractable derivatives have expanded the available strains for experiments. But even still, these laboratory strains represent a small fraction of the diversity of yeast. The continued development of new laboratory strains will broaden the potential questions that can be posed. We are now poised to take advantage of this diversity, rather than viewing it as a detriment to controlled experiments.
Assuntos
Evolução Molecular , Genética Microbiana/métodos , Técnicas Microbiológicas/métodos , Biologia Molecular/métodos , Saccharomyces cerevisiae/genética , Genética Microbiana/história , História do Século XX , História do Século XXI , Técnicas Microbiológicas/história , Biologia Molecular/históriaRESUMO
The budding yeast Saccharomyces cerevisiae is an outstanding experimental model organism that has been exploited since the early part of the twentieth century for studies in biochemistry and genetics. It has been the premiere experimental system for modern functional genomics and continues to make important contributions to many areas of biology. Here we discuss its many virtues as an organism for classical genetic research.
