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Lesson 9 Flashcards

Cytogenic techniques, Genetic Engineering, & Gene Therapy

1
Q

The study of chromosomes using traditional cytogenetic techniques requires cells that are actively dividing

A

Cytogenetic Technique

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2
Q

Chromosomes are individually distinguishable under _____ microscope only during cell division and are best examined during ________

A

Light microscope; metaphase

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3
Q

Metaphase chromosomes can be obtained from specimens that contain

A

spontaneously dividing cells
cultured
and chemically induced to divide in vitro

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4
Q

Specimens that contain spontaneously proliferating cells include:

A
  • bone marrow
  • lymph nodes
  • solid tumors
  • chorionic villi
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5
Q

What are routinely cultured to obtain dividing cells

A

Peripheral blood lymphocytes
tissue biopsies
amniotic fluid samples

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6
Q

Lymphocytes usually require

A

The additional of a mitotic stimulant

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7
Q

It depends on clinical indications and whether the diagnosis is prenatal or postnatal

A

The choice of specimen for chromosome analysis

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8
Q

The most critical requirement of culture initiation, maintenance, and cell harvest

A

Living cells capable of cell division must be received by the laboratory

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9
Q

The laboratory may reject specimens that are

A

improperly labeled or unlabeled

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10
Q

Living cells
Sterility
Proper growth medium
+/- Mitotic stimulant
Microbial inhibitors

A

Culture Initiation

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11
Q

Sterility
Optimal temperature
Optimal pH
Optimal humidity
Optimal time interval

A

Culture Maintenance

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12
Q

Arrest division
Swell cells
Fix cells
Prepare slide
Stain/band

A

Cell Harvest

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13
Q

2 broad categories of Chromosome banding and staining

A
  1. Produce specific alternating bands along the length of each entire chromosome
  2. Stain only a specific region of some or all chromosomes
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14
Q

This creates unique patterns for positive identification and permits characterization of structural abnormalities; numerical and structural examination

A

Specific alternating bands

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15
Q

Used in special circumstances when a particular piece of information cannot be answered using a routine banding method

A

Stain only a specific region

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16
Q

Techniques that create bands along the length of the chromosomes

A

G-banding
Q-banding
R-banding

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17
Q

Techniques that stain selective chromosome regions

A

C-banding
T-banding
Cd-banding
G-11 banding
NOR
DAPI/DA Staining
FISH

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18
Q

G bands produced with trypsin and Giemsa

A

GTG banding

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19
Q

dark bands in G-banding

A

A-T rich (late replicating, heterochromatic regions)

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20
Q

light bands in G-banding

A

C-G rich (early replicating, euchromatic regions)

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21
Q

The biologically more significant ACTIVE REGIONS

A

G-light bands

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22
Q

The relatively few active genes

A

G-dark bands

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23
Q

other stains used in G-banding techniques

A

(Giemsa)
Wright’s
Leshman’s

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24
Q

Fluorescent technique that was first developed for human chromosomes

A

Q-banding

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25
What certain fluorochromes will bind to DNA and produce distinct banding patterns of bright and dull fluorescence when excited with the proper wavelength of light
quinacrine dihydrochloride
26
The brightly fluorescing regions are what in Q-banding
A-T rich
27
What differentiates Q-banding from G-banding?
distal long arm of the Y fluoresce brightly
28
Produce a banding pattern that is the OPPOSITE OR REVERSE of the G-banding pattern
R-banding
29
Methods in R-banding
fluorescent nonfluorescent
30
R-banding C-G rich
heterochromatic regions that stain darkly (bright)
31
R-banding A-T rich
euchromatic regions that stain lightly (dull)
32
What is the positive region in R-banding?
light stained, fluoresce dull, A-T rich euchromatic
33
C-banding means
Constitutive Heterochromatin Banding
34
Where does C-banding selectively stain?
the constitutive heterochromatin around the centromeres
35
Useful for determining the presence of dicentric and pseudodicentric chromosomes = marker chromosomes and polymorphic variants
C-banding
36
Resists degradation
Constitutive heterochromatin
37
Selectively depurinated and denatured by barium hydroxide
CBG banding
38
Results in only the terminal ends or TELOMERES
T-banding
39
Pair of dots at each centromere, one on each chromatid that differentiate functional from nonfunctional centromeres
Cd staining
40
Stains the pericentromeric regions of all chromosomes at high alkaline pH to delineate heterochromatin polymorphisms
G-11 banding
41
Selectively stains silver nitrate on the nucleolar organizer regions located on the satellites stalks of the acrocentric chromosomes
NOR
42
Theoretically, how many NORs are per cell and, for each acrocentric chromosome?
10 NORs, 1 for each acrocentric chromosome
43
Combines DAPI (fluorescent dye) w/ distamycin A (nonfluoro antibiotic) to certain A-T rich areas of constitutive heterochromatin in C-band regions for differentiating between satellite regions of any of the acrocentric chromosomes (1,9,16 distal, short arm 15)
DAPI/DA staining (4,6-diamino-2phenole-indole/ distamycin A)
44
Uses fluorescent probes that attaches to specific areas in the chromosome with high degree of sequence complementarity
FISH
45
Used to detect and localize presence or absence of specific DNA sequences in the chromosomes to determine the possible cause of a child's developmental disability
FISH
46
Collection of microscopic DNA spots (traditional solid/alt bead array)
DNA microarray analysis
47
Manipulation of an organism's nucleic acids or the insertion of DNA into cells
Genetic engineering
48
One of whose genes have been artificially manipulated for a desired effect
Genetically Modified Organisms (GMO)
49
Technology used to cut a known DNA sequence from one organism and introduce it into another organism thereby altering the genotype and therefore altering the phenotype of the recipient
Recombinant DNA technology
50
Process of isolating genes from one organism, manipulating purified DNA in vitro, and transferring to another organism
Gene cloning
51
Who started gene cloning when discovering enzymes degrading bacterial viruses
Arber (1950)
52
Who purified enzymes and characterized them to be able to cut DNA at specific sites?
Smith (1970)
53
Purified enzymes that recognizes a specific sequence of bases and cut the DNA backbone
Restriction enzymes (restriction endonucleases)
54
Recombinant vectors
-plasmids -specially designed bacteriophage lambda chromosomes -cosmids
55
the cut DNA is introduced into
the plasmid
56
Restriction enzymes acts as endonucleases to cleave a DNA sequence at a specific site called
Restriction site (recognition sequence)
57
Plasmid containing the cloned gene
Chimera
58
The DNA sequence that has been incorporated in the vector
Insert
59
Methods of inserting DNA into a cell
- heat shock method - electroporation - genetically engineered viruses - gene gun - microinjection - liposomes
60
When chimera plasmids are placed in a solution (w/ cold CaCl2 + normal host bacteria) with ABRUPT HEATING at 42°C for 2-5 minutes
Heat shock method
61
When host cells are exposed to HIGH VOLTAGE PULSE
Electroporation
62
Genetic engineering applications
- therapeutic applications - gene therapy - chimeric monoclonal antibodies - HGP/ Human Proteome Project - Scientific applications
63
Experimental technique that utilizes genes to treat or prevent diseases by inserting a gene into a patient's cells instead of using medicines/drugs or surgery
Gene therapy
64
Two main types of gene therapy
1. Somatic cell 2. Reproductive or germ-line
65
Genes are introduced in an organ or tissue to induce production of specific products DOES NOT AFFECT the individual's GENETIC MAKEUP as a whole (transmitted to the next) Allows damaged organs to function properly
Somatic cell gene therapy
66
Integration of corrective genes into sex cells or zygotes with the goal of generating beneficial genetic change that CAN BE TRANSMITTED to the progeny Offspring can inherit the introduced genes
Reproductive or germ-line gene therapy
67
Medical conditions for which gene therapy is being investigated on:
- bubble boy disease - hemophilia - AIDS - cancer (liver, lung. breast, colon, ovarian, prostate) - asthma - brain tumor - melanoma - muscular dystrophy - neurodegenerative conditions - diabetes - heart diseases

Cytogenetics (11 decks)

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