Genome manipulation techniques
13 important questions on Genome manipulation techniques
What are two methods for forward genetics?
- Transposons
What are four methods for reverse genetics?
- Transgenesis
- Homologous gene targeting
- CRISPR/Cas9
What is the goal of mutagenesis and the use of transposons?
- Higher grades + faster learning
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How are transposons used in fruit flies?
How does transgenesis work?
- which randomly integrates somewhere in the genome,
- which leads to overexpression of the gene (which is often a mutant gene)
What are the parts of a transgenic construct?
- (An intron (with regulatory elements))
- Transgene (cDNA)
- Poly(A) signal
What are the two endogenous genome editing methods?
- CRISPR/Cas9
What are the difference between NHEJ and HR?
How does homologous gene targeting work?
What does the targeting vector contain?
- A selectable marker in the middle (which can be a null allele, mutant allele or a floxed (conditional) allele)
How is homologous gene targeting used in mice?
2. The ES cells are injected in the blastocyst, which produces chimeras
3. A mouse colony is breeded
What is the difference between transgenesis and homologous gene targeting as regards the cell stage in which the injection (HGT: ES cells, T: DNA) is placed?
Transgenesis: oocytes
What is a disadvantage of homologous gene targeting?
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- Never study anything twice again
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