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Cytogenetics > Lesson 9 > Flashcards

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
Q

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

A

quinacrine dihydrochloride

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26
Q

The brightly fluorescing regions are what in Q-banding

A

A-T rich

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27
Q

What differentiates Q-banding from G-banding?

A

distal long arm of the Y fluoresce brightly

28
Q

Produce a banding pattern that is the OPPOSITE OR REVERSE of the G-banding pattern

A

R-banding

29
Q

Methods in R-banding

A

fluorescent
nonfluorescent

30
Q

R-banding C-G rich

A

heterochromatic regions that stain darkly (bright)

31
Q

R-banding A-T rich

A

euchromatic regions that stain lightly (dull)

32
Q

What is the positive region in R-banding?

A

light stained, fluoresce dull, A-T rich euchromatic

33
Q

C-banding means

A

Constitutive Heterochromatin Banding

34
Q

Where does C-banding selectively stain?

A

the constitutive heterochromatin around the centromeres

35
Q

Useful for determining the presence of dicentric and pseudodicentric chromosomes = marker chromosomes and polymorphic variants

A

C-banding

36
Q

Resists degradation

A

Constitutive heterochromatin

37
Q

Selectively depurinated and denatured by barium hydroxide

A

CBG banding

38
Q

Results in only the terminal ends or TELOMERES

A

T-banding

39
Q

Pair of dots at each centromere, one on each chromatid that differentiate functional from nonfunctional centromeres

A

Cd staining

40
Q

Stains the pericentromeric regions of all chromosomes at high alkaline pH to delineate heterochromatin polymorphisms

A

G-11 banding

41
Q

Selectively stains silver nitrate on the nucleolar organizer regions located on the satellites stalks of the acrocentric chromosomes

A

NOR

42
Q

Theoretically, how many NORs are per cell and, for each acrocentric chromosome?

A

10 NORs, 1 for each acrocentric chromosome

43
Q

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)

A

DAPI/DA staining

(4,6-diamino-2phenole-indole/ distamycin A)

44
Q

Uses fluorescent probes that attaches to specific areas in the chromosome with high degree of sequence complementarity

A

FISH

45
Q

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

A

FISH

46
Q

Collection of microscopic DNA spots (traditional solid/alt bead array)

A

DNA microarray analysis

47
Q

Manipulation of an organism’s nucleic acids or the insertion of DNA into cells

A

Genetic engineering

48
Q

One of whose genes have been artificially manipulated for a desired effect

A

Genetically Modified Organisms (GMO)

49
Q

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

A

Recombinant DNA technology

50
Q

Process of isolating genes from one organism, manipulating purified DNA in vitro, and transferring to another organism

A

Gene cloning

51
Q

Who started gene cloning when discovering enzymes degrading bacterial viruses

A

Arber (1950)

52
Q

Who purified enzymes and characterized them to be able to cut DNA at specific sites?

A

Smith (1970)

53
Q

Purified enzymes that recognizes a specific sequence of bases and cut the DNA backbone

A

Restriction enzymes (restriction endonucleases)

54
Q

Recombinant vectors

A

-plasmids
-specially designed bacteriophage lambda chromosomes
-cosmids

55
Q

the cut DNA is introduced into

A

the plasmid

56
Q

Restriction enzymes acts as endonucleases to cleave a DNA sequence at a specific site called

A

Restriction site (recognition sequence)

57
Q

Plasmid containing the cloned gene

A

Chimera

58
Q

The DNA sequence that has been incorporated in the vector

A

Insert

59
Q

Methods of inserting DNA into a cell

A
  • heat shock method
  • electroporation
  • genetically engineered viruses
  • gene gun
  • microinjection
  • liposomes
60
Q

When chimera plasmids are placed in a solution (w/ cold CaCl2 + normal host bacteria) with ABRUPT HEATING at 42°C for 2-5 minutes

A

Heat shock method

61
Q

When host cells are exposed to HIGH VOLTAGE PULSE

A

Electroporation

62
Q

Genetic engineering applications

A
  • therapeutic applications
  • gene therapy
  • chimeric monoclonal antibodies
  • HGP/ Human Proteome Project
  • Scientific applications
63
Q

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

A

Gene therapy

64
Q

Two main types of gene therapy

A
  1. Somatic cell
  2. Reproductive or germ-line
65
Q

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

A

Somatic cell gene therapy

66
Q

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

A

Reproductive or germ-line gene therapy

67
Q

Medical conditions for which gene therapy is being investigated on:

A
  • 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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