LECTURE XI Flashcards
why genetic test?
prenatal diagnosis, heterozygote carrier detection, presymptomatic diagnosis of genetic disease
how to genetic test?
analysis
treatment
disease management
what are some examples of genome sequencing technology?
applied biosystem biosystems
Oxford nanopore minion
components of informed consent?
minimize risk to subjects sound research when appropriate risks are reasonable risks vs benefits weighed long range effects special problems informed consent monitor data collected patient privacy
what are the elements that comprise HIPAA?
core elements
required elements
optional elements
what are the core elements of HIPAA?
name other persons involved description of purpose authorization expiration date signature of individual
what are the genetic techniques?
nucleic acid visualization
PCR
RFLP
DNA sequencing
what are the mutation detection techniques?
RFLP
allele specific Oligos
DNA sequencing and mutation detection
trinucleotide
what are the different levels of genetic testing?
DNA
Protein
Protein Function
what kind of genetic testing is used for large changes in chromosomes, extra chromosomes, very large deletions or insertions?
analysis of whole chromosomes at the DNA level
what kind of genetic testing is used for small changes; mutations in the sequence, small deletions or insertions?
analysis of sequence at the DNA level
what kind of genetic testing is used for any change that may affect the folding of the protein?
analysis of protein shape at the protein level
what kind of genetic testing is used for the functional protein?
analysis of protein function at the protein level
example of single base pair mutation?
sickle cell anemia
example of deletion?
cystic fibrosis
Duchenne muscular dystrophy
example of insertion?
huntingtons disease
example of multiple mutations?
diabetes, susceptibility to breast cancer
how do we amplify target sequence?
polymerase chain reaction
melting annealing elongation melting annealing elongation
thus PCR test for deletion uses electrophoresis
this type of analysis looks for polymorphism between people in the number of restriction sites?
RFLP analysis
what are the issues of RFLOP analysis?
can not detect all mutations, the mutation must coincide with a restriction enzyme cut site
what are the issues with sequencing?
informative
time consuming
expensive
when is a dot blot used?
querying DNA or PCR products using radioactive oligomers; can be used to detect CF
what type of analysis do we use to find out if patients DNA sequence is different?
sequencing
this allows for the testing of many different mutations via many high throughput dot blots?
microarray
as a result of clinical validity, what is the significance of DNA based tests?
genetic testing genetic screening -heterozygote screening -newborn screening -prenatal diagnositics -presymptomatic testing
what are the many techniques for prenatal genetic testing?
chronic villus sampling or amniocentesis
biomarker assays
ultrasonography
direct DNA based test
this is a test used for couples at significantly increased risk for transmitting a harmful or predisposing allele to offspring?
preimplantation genetic diagnosis (PGD)
what is the criteria for ideal candidates for gene therapy?
single defect
well characterized
significant morbidity and mortality
cells are experimentally accessible
what do the genes into cells rely on?
viral or nonverbal vector system
this is the introduction of a gene, or DNA sequence, into a cell to correct a genetic defect?
gene therapy
this involves the alteration of genes in human somatic cells to treat a specific disorder?
somatic cell therapy
this is a type of RNA virus that can insert its genome into the host cell after reverse transcribing their viral RNA into dsDNA (what is this called)
what is the disadvantage?
retroviral vector
transduction
due to preferentially integrating near promotor it can add into a proto-oncogene activating tumorogenesis getting into the nucleus when dividing
this is due to inability of retroviruses to add into slowly or non dividing cells, other delivery systems instituted. Virus can be used in vaccines. Can accept inserts as large as 36kB in size
what is a disadvantage?
adenoviral vectors
DNA is not integrated into a chromosome and so it will not activate the proto oncogene and so transient expression results. This vector provokes an immune response
this is a complex of RNA retroviruses that can also transduce non dividing cells through nuclear membrane, similar to HIV. This can accept inserts of 8kB. This type of vector is considered a focus of gene therapy protocols?
lentiviral vector
challenges of viral gene therapy?
transient low level expression
difficulties reaching specified tissue/cells
necessity for precise regulation of gene activity
potential for insertion mutagenesis
gene blocking/silencing therapies?
antisense oligonucleotides
ribozymes
RNAi
