Transgenic Organisms Flashcards
What are micro injections
Direct delivery of DNA into nucleus with a microcapillary needle and micromanipulator.
This is most common to generate transgenic organisms like mice as it has precise control but because of that it is very time consuming and challenging.
What are biolistics
DNA is coated onto gold or tungsten particles and fire into cells at high pressure using helium (gene gun).
This is used when creating transgenic plants when it’s hard to transfect target cells or tissues. But problem is DNA is inserted randomly so little control.
What is electroporation
Applies an electrical field to cells increasing the permeability of cell membrane allowing insertion of DNA.
This has high transfection efficiency and versatility. But may cause substantial cell damage leading to death.
How does agrobacterium mediated transformation work
Agrobacterium component cells are co-cultivated with plant tissues.
Agrobacterium transfers T DNA of interest from its Ti plasmid into the plant cell which integrates into plant genome and then specific trait is passed onto next generation.
The T-DNA can be modified using restriction enzymes and ligases to insert DNA of interest (up to 30 kB)
How TALENS are used in gene editing
Pair of DNA binding proteins fused to Fok1 nuclease.
They recognise the target site based on DNA protein interactions allowing for high specificity and precision with low cytotoxicity. Also cost effective and easy to use.
How does CRISPR work
RNA guided endonuclease cas9 and single guided RNA that recognises target sequence.
DNA site is recognised by complementary bases between genomic DNA and RNA guide and is loaded into cas9 nucleases cleaving the DNA.
What are Difference between gene editing and other transformation methods
Gene editing tech is more precise modification of genome (gene knockout, fixing genetic defects)
Other transformation methods introduces foreign DNA but not as precise
