RR7 Flashcards
If transcription does not occur in a linear manner, how does it occur?
It is 3-dimensional with topographical changes within DNA and chromatin.
How does you make a specific transcript?
Decorate RNA to quantify how much is produced. This introduces elements into RNA so that it folds into a 2nd structure that is recognized by specific proteins. Attaches to GFP. Now you can introduce trans gene, one to be transcribed and one with GFP protein which flags gene, it has the ability to interact gene with specific cRNA structure. If the trans gene is properly expressed, then at the 5’ end of RNA there will be a structure recognized by GFP that can bind.
How does the embryo develop in the fly?
All cell membranes are marked with GFP type protein. After nuclear divison, there is a signal that tells cells to migrate inwards, this is when morphology shows.
What are shadow enhancers?
Shadow enhancers have the potential to evolve novel binding sites and achieve new regulatory activities without disrupting the core patterning functions of critical developmental control genes. They can either take enhancers and put it at the 5’ transcriptional start site or on the 3’ site. Or they can not put it at all.
When RNA pol ll generates an RNA molecule, what will we see?
We will see small green dots at the 3’ or 5’ end.
What does transcription efficiency correlate with?
It correlates with increased “bursts” frequency. This is the frequency of burst that take place from a given promoter that is being activated.
How can burst frequency be changed?
It can be changed with respect to the strength of the enhancer. In a weak enhancer, burst frequency is reduced.
Which burst more: highly transcribed promoters or low transcribed promoters?
Highly transcribed promoters
Do p-granules go to the posterior end or anterior? Why?
They go to the posterior end because of chemical compposition.
Do p-granules go to the posterior end or anterior? Why?
They go to the posterior end because of chemical composition.
What are p-granules? How were they discovered?
P-granules are “liquid-liquid” condensates that contain RNA molecules. GFP labelling of mediator suggest that it forms large complex liquid-like structures in cells, they are large clumps that contain med-I GFP than a single molecule.
How do you mediate liquid-liquid phase seperations?
They use intrinsically disordered domains. mCherry - see agglomerations within nuclei.
What is an intrinsically disordered domain?
It is a domain that we can’t obtain the structural information of.
Describe the formation of liquid-liquid condensates:
