RR7

Question 1

How can we mark specific transcripts?

Answer 1

1) Introduce elements into RNA so that it folds into a 2nd structure that is recognized by specific proteins - attach to GFP
2) Introduce 2 things to transgene - one to be transcribed, one with GFP

If the transgene is properlyn expressed, then at the 5’ end of RNA there will be a structure recognized by GFP that can bind

This is decorating RNA to quantify how much is produced

Question 2

Describe how we mark the embryo development in the fly

Answer 2

Mark the cell membranes with GFP type protein
- After nuclear division, the development signal tells cells to migrate inwards
- This is when morphology shows

Question 3

What is a shadow enhancer?

Answer 3

• An enhancer critical to expression
• Can be put on 3’ end or 5’ end or not at all
  We can take image of cells - will see small green dot enhancing the shadow enhancer - allows us to determine 3’ or 5’ side

Question 4

What is “burst” frequency?

Answer 4

Increased efficiency of RNA synthesis is based on frequency of bursts that take place from given promoter that is being activated
Frequency changes with respect to enhancer strength
Weak enhancer means reduced burst frequency

Question 5

What are P-granules?

Answer 5

Liquid-liquid condensates that contain RNA molecules
Go to posterior or anterior end based on concentration of proteins in specific regions

Question 6

How are liquid-liquid condensates formed?

Answer 6

Formed by transcriptional regulators
Ex: a bromodomain

Question 7

What impacts the formation of liquid liquid condensates?

Answer 7

• Concentration (DNA, RNA, protein)
• Valency
  Mediator and RNA pol II form liquid-like structures in cell during transcription