Offner Techniques in Molecular Medicine

Question 1

What is the one requirement of all restriction sites

Answer 1

They have to be palindromes
Ex. GAATTC

Question 2

RFLP

Answer 2

1) cut DNA with RE
2) Gel, membrane, denature DNA
3) ssDNA incubated with probes
4) X-ray indicates presence of either fragment

Question 3

What are STRs (short tandem repeats)?

Answer 3

2-6 bases repeated a variable number of times
- occurs in the same part of the genome for all individuals
- conserved in 13 spots in the genome
- replicated by PCR to identify people

Question 4

CRISPR Limitations

Answer 4

• delivery system
• off target cleavage/effects
• low knock-in efficiency
• ethics

Question 5

Knock-out (NHEJ)

Answer 5

Create sgRNA to guide Cas9 to cutting site
- 2 ends rejoin and have in/del

Question 6

Knock-In (HR)

Answer 6

Cut and paste sequence
- very inefficient

Question 7

How is sickle cell treated using CRISPR

Answer 7

BCL knocked out allow expression of gamma-globin

Question 8

How is wet AMD treated

Answer 8

Injection of anti-vEGF into eyes to control growth of blood vessels

Question 9

How can lung cancer be treated

Answer 9

inhalation of siRNA nanoparticles