Techniques

Question 1

Immunoprecipitation

Answer 1

• It is the technique of precipitating a protein antigen out of solution by means of an antibody that specifically binds to that particular protein
• This process can used to separate and isolate a protein out of a solution containing thousands of proteins
• This technique was used in the YAP/TAZ paper to precipitate out these proteins, and see what was attached to YAP and TAZ

Question 2

Western Blotting

Answer 2

• A technique that separates a protein mixture based on the size of the polypeptide
• Makes use of antibodies to highlight the protein of interest

Question 3

Genetic reporting

Answer 3

• A reporter gene is a gene that researchers attach to a regulatory sequence of another gene of interest in bacteria, cell culture, etc
• Reporters have the same promoters as a target TF
• Used to measure activity of a transcription factor

Question 4

Immunofluorescence

Answer 4

• Use of a primary antibody to find protein antigen: secondary antibody conjugated to immunofluorescence
• We can observe the activity of specific proteins in the context of a cell

Question 5

siRNA knockout

Answer 5

• The technique by which the RNA for a certain protein is not expressed, leading to inhibited function that protein is responsible for

Question 6

Electron microscopy

Answer 6

• Way to visualize the shape of sub-cellular structures like proteins

Question 7

Transposon flanked expression cassettes

Answer 7

The way that they viewed the SHH going up the filopodia