Examples Viral transduction into mammalian cells Agrobacterium transformation of plant cells Sources httpenwikipediaorgwikiViralvector Daniel Eisenman Medical University of South Carolina ID: 1045599
Download Presentation The PPT/PDF document "Chapter 17: Introduction of DNA into euk..." is the property of its rightful owner. Permission is granted to download and print the materials on this web site for personal, non-commercial use only, and to display it on your personal computer provided you do not modify the materials and that you retain all copyright notices contained in the materials. By downloading content from our website, you accept the terms of this agreement.
1. Chapter 17: Introduction of DNA into eukaryotic cells – biological methodsExamples:Viral transduction into mammalian cellsAgrobacterium transformation of plant cellsSources: http://en.wikipedia.org/wiki/Viral_vectorDaniel Eisenman, Medical University of South CarolinaNemat O. Keyhani; cited papersinternet1
2. OutcomesExplain how the procedures described workDistinguish the types of viral vectors used to introduce DNA into cellsDescribe and explain how safety is maintained when working with viral vectorsDescribe and explain how the Agrobacterium system is modified for introducing DNA into plant cells2
3. Viral vectors for transduction (transfer) of DNA into mammalian cells are disarmed:Genes required for disease have been removed (but the viral coat proteins can still cause immuno-responses)Ability to propagate is usually also removed3
4. Viral vectors for transduction (transfer) of DNA into mammalian cellsAdvantage (compared to e.g. lipofection) : Nearly all virus particles infect cellsTherefore suitable for: High production of proteinRNA interferenceGene therapyDisadvantage: It is complicated to generate suitable vectors and virus particlesCompanies provide kits that simplify the cloning and transfection procedure (for $$$)4
5. Types of viral vectorsRetrovirusesRNA virusesCan integrate into genomes, but only in dividing cellsLentivirusesRNA virusesCan integrate into genomes, ALSO in non-dividing cellsAdenovirusesDNA virusesCan’t integrate into genomeMay cause rapid immune responseAdeno-associated virusDNA virusCan’t integrate into genomeUsually don’t infect mammalian cells weak/no immune response5
6. Retro- and LentivirusesSingle stranded RNA genomeHost range determined by envelope proteinsHave a mechanism for converting RNA into DNA and integrating it into host genomeHost Cellhttp://www.biology-pages.info/R/Retroviruses.html6