Modern Technologies

Question 1

Microarrays

Answer 1

Small glass or silicon slides featuring oligonucleotides

Use hybiridization

Use fluoresence scanner

High-throughput process

Question 2

What can microarrays do>

Answer 2

Expression profiling - determine what genes are active in a population of cells and at what levels (compare normal cells vs tumors)

SNP profiling - ID SNPs associated with certain diseases (compare individuals within population)

Question 3

Expression profiling process

Answer 3

Create cDNA from RNA…cDNA will attach to oligonucleotides and can be measured via fluorescence (attached to cDNA)

Question 4

Expression profiling steps

Answer 4

Prepare mRNA, generate cDNA, hybridize cDNA to microarray, analyze data (how much cDNA hybridized to each probe?)

Question 5

Higher expression/lower expression in cancer on Microarray

Answer 5

Higher - oncogenes

Lower - tumor suppressor genes

Question 6

What can microarrays NOT do?

Answer 6

Meausre changes at translational or post-translational level

Question 7

SNP analysis

Answer 7

Immobile sequence hybridize SNPs at a particular location…can determine homo or heterozygous

Question 8

Microarrays evaluate differences in levels of

Answer 8

Numerous mRNA (entire transcriptome) between cell populations OR

Monitor SNP differences between individuals in a population

Question 9

High Throughput sequencing

Answer 9

Sequence short amounts at a time

Use adapter at the ends and sequence inward

Line up region with reference genome sequence and determine if hetero or homozygous for a polymorphism

Question 10

RNA -seq analysis

Answer 10

mRNA converted to cDNA…RNA sequences will align with exons

Useful for detecting splice variants

Question 11

CRISPR RNA - Guided nucleases

Answer 11

Clustered regularly interspaced short palindromic repeats

Can bind specific sequences of DNA and cleave in a very specific location

Can be used to mutate DNA or potentially repair a mutated gene

Question 12

iPS are

Answer 12

Induced pluripotent stem cells…can be formed from 4 specific transcription factors

Question 13

iPS cells can differentiate

Answer 13

in vitro

Question 14

Application of iPS cells

Answer 14

Can take out patient’s cellsm reprogram them, correct gene using CRISPR RNA, allow differentiation, and transplant back into the patient

Question 15

What is essential for GWAS studies

Answer 15

SNP microarray usage