genomics- Stem Cells (unit 3) Flashcards
goals of HGP
- DNA based genetic testing for disease and carrier status
- pharmacogenetics for new drugs
shotgun sequencing
1) isolate DNA, sequence lots of it
2) use computer to line up short DNA sequences
genetics vs genomics
genetics= single DNA molecule to one phenotype genomics= multiple molecules- whole genome to metabolomics
transcriptomics
the genes expressed in a given cell type or environmental condition =analysis of transcripts- level of mRNA expressed
-uses micro assay and RNA sequencing
DNA microassay
high-throughput tech used in molecular bio and in medicine- allow the expression level of thousands of genes to be measured at the same time
=arrayed series of 1000s of microscopic DNA, each containing specific DNA sequence
Reverse transcriptase
enzyme that makes DNA copy (cDNA) of RNA molecule
cDNA
copy of mRNA
proteomics
large-scale analysis of proteins of the proteome; all the proteins in a cell or tissue
methods= immunoprecipitation or mass spec
interactome
which proteins interact (bind/ regulate expression) with which other proteins
protein networks
pairs of interacting proteins= identified by Y2H and other methods, can be linked together in networks
synthetic bio
using engineering principles and bio components to purposefully make up and modify organisms
biobricks
promoters, enhancers, operators, repressors, terminators, protein domains, enzymes, siRNAs, miRNAs, etc…
reporter genes
investigate gene expression patterns
LacZ with B gal
GFP used to label specific cells
chimeras
animals composed of cells from different genetic sources
microinjecting DNA
most common method used to produce transgenic farm animals (into fertilized egg)
pharming
using transgenic animals to produce drug compounds
somatic cell nuclear transfer
cloning method also used to add transgenes
transgenic plant methods (2)
agrobacterium (uses gene plasmid into bacteria and then mix with plant cell, then plasmid will mix with chrom, and then screen for cells with trans gene) gene gun (gold particles coated with DNA are shot at cell with gene gun and DNA incorporated into plant cell chromosome to screen for cells with transgene
Bt toxin
plant (transgenic) makes Bt toxin in inactive form, insect eats, insect converts to active form, and then dies… protecting future plant progeny
CRISPR
clustered regularly interspaced short palindromic repeats
-natural role= bacterial acquired immunity
immunization= 1) phage DNA sampled and 2) spacers are integrated between repeats = proteins incorporate phage DNA at Eds of repeats- recognizable
PAM
must be right upstream of match (N-GG)
natural role
1) bacteria have ability to acquire long-term immunity to threats they experience
2) widely spread through bacteria
3) many variants with differing proteins and mechanisms of action
4) similarities to RNAi suggest lab modification potential
RNA guided nuclease
nucleic acid can direct a nuclease to almost any sequence in any genome- and being able to break DNA at specific locations gives the ability to change DNA at any location
-allows creation of a DBS by Cas9 at almost any location which can be repaired by NHEJ or HDR
non homologous end joining (NHEJ)
mistakes are made- insertions and deletions randomly occur
homology dependent repair (HDR)
depends on source of homology sequence
-if HDR occurs over NHEJ, if a truly homologous sequence is used, then it will be cut again and again until NHEJ causes indel