Gene Silencing

Question 1

What is RNA interfernece?

Answer 1

Accurate and potent gene silencing method that silences RNA by breaking it down or stopping translation

dsRNA

Question 2

What are the types of siRNAs?

Answer 2

Question 3

What causes RNAi?

Answer 3

siRNA

Question 4

What does RNAi do?

Answer 4

Promote the degredation of mRNA

Question 5

Where does dsRNA eom from?

Answer 5

Question 6

What is DiceR?

Answer 6

Processes dsRNA and converts to smaller dsRNA molecules

Question 7

What is the short dsRNA called?

Answer 7

siRNA which has 21-23 nucleotides with 3’ two nucleotide overhang

Question 8

When siRNA interacts with dicer what does it activate?

Answer 8

RISC which cleaves the passenger strand of the siRNA while the guide strand remaines associate with the RISC

Question 9

What does the guide strand do?

Answer 9

Activate the RIS to target its target mRNA for cleavage by AGO2

Question 10

What genes does the guide strand bind to?

Answer 10

Only binds to strands that is fully complementary to it siRNA causing specific gene silencing

Question 11

***REVIEW THE NEXT IMAGE!

Answer 11

Question 12

The RISC and AGO complex join together and use the siRNA strand to?

Answer 12

Bind to the complimentary mRNA for cleavage

Question 13

What can the argonaute protein do when bound to the complimentary strand?

Answer 13

Recruit accerssory factors to regulate the target sequence in other ways

Question 14

What is miRNA?

Answer 14

Class of moleucles that negatively regulate gene expression

Question 15

What is the difference between miRNA and sirna?

Answer 15

the sequence does not need to be fully complimentary to the compared strand

Question 16

What transports the miRNA out of the nucleus and into the cytoplasm?

Answer 16

exportin5

Question 17

What does DICER do?

Answer 17

Cleaves the ends of the miRNA and siRNA making it compatible with the RIS-AGO enzyme

Question 18

What is a difference between miRNA and SiRNA in terms of the risc-ago2 complex?

Answer 18

Question 19

What causes the discarding of the miRNA strand/

Answer 19

The passenger strand is unwound from the miRNA duplex leading to it being discarded

Question 20

What is the major difference between siRNA and miRNA?

Answer 20

Question 21

What is the benefit of the miRNA and mRNA recognition?

Answer 21

One miRNA strand can recognize an array of mRNAs and hence miRNA has the charactertistics of having multiple target

Question 22

What is the benefit of RNAi therapeutics?

Answer 22

Question 23

Review the next chart

Answer 23

Question 24

What are the limitations of RNAi?

Answer 24

Vulnerable to serum nucleases, degraded rapidly, short half-life, poor stability.

Question 25

What is the role of delivering RNAi?

Answer 25

Facilitate the cellular uptake of siRNA and miRNA to their target sites, protext the nucleic acids from premature nuclease degredation

Question 26

What are some delivery systems for RNAi?

Answer 26

Viral vectors, polymer based and lipid based systems

Question 27

Review next slide

Answer 27

Question 28

What does crispr components make up?

Answer 28

Guide RNAs, cas9, DNA templates

Question 29

What is the BCL11A gene?

Answer 29

This repressed the fetal hemoglobin expression hence, by turning this off we can start producing fetal hemoglobin again and fix sickle cell anemia