Optogenetics: Exam 2

Question 1

Optogenetics

Answer 1

expressing light sensitive proteins in neurons, turn on function with light: manipulating gene expression to alter neuronal activity in behaving animals

Question 2

Are genes expressed?

Answer 2

amount mRNA/protein, fluorescence microscopy etc.

Question 3

Fluorescence Microscopy

Answer 3

detect proteins: fluorophores (fluorescent molecules): high energy/short wavelength/blue light –> lower energy/red light is emitted, filters used to isolate excitation/emission wavelengths

Question 4

Immunofluorescence

Answer 4

Technique to make structure fluorescent: using fluorophore-tagged antibodies to target specific proteins

Question 5

Green Fluorescent Protein

Answer 5

Technique to make structure fluorescent: insert gene for GFP into genome of cell

Question 6

Transgene

Answer 6

gene transferred from genome of one organism into another (external gene)

Question 7

Transgenic

Answer 7

genetically engineered cell/organism generated by inserting transgene into host cell

Question 8

GFP fusion protein

Answer 8

examine protein localization in cell

Question 9

Immunoblots

Answer 9

examine presence of protein by using antibodies that bind to proteins selectively

Question 10

DNA Microarrays

Answer 10

visualize “transcriptome” at any time/particular condition: “chips” contain wells

Question 11

Transcriptome

Answer 11

what genes are expressed in a cell at time when experiment is done; advantage: high throughput, disadvantage: only shows transcribed potentially not translated

Question 12

RT (reverse transcription)- PCR (4 steps)

Answer 12

detects presence of mRNA: 1. isolate mRNA, 2. use reverse transcriptase to ‘reverse transcribe’ mRNA -> DNA, 3. PCR to amplify DNA segments, 4. gel electrophoresis

Question 13

Recombinant DNA

Answer 13

formed artificially by combining DNA segments of different origins: relies on restriction enzymes, DNA vectors

Question 14

Channelrhodopsin (ChR)

Answer 14

techniques used to genetically engineer neurons to epress light-sensitive gene that codes for protein ChR

Question 15

Restriction Enyzymes

Answer 15

Cut DNA in specific location, found in bacteria, palindromes (read same back and forth) Check photo on DT

Question 16

DNA Vectors

Answer 16

Often circular forms of DNA, carry/replicate/express DNA segments of interest:
Can use restriction enzyme to open vector so another DNA segment can be inserted:
Can gene –> vector –> target cell

Question 17

Adenoviruses

Answer 17

used to deliver vectors with gene of interest into target cell

Question 18

CRISPR - Cas9 system

Answer 18

Insert genes into target genomes: guide RNA base pairs with target region of SNA, Cas9 enzyme cleaves DNA sequence at specific site: can alter genes that effect how neurons function