Genetics and more biology Flashcards
(77 cards)
1
Q
BR
A
C57BR strains
2
Q
HR
A
HRS strains
3
Q
L
A
C57L strains
4
Q
R3
A
RIIIS strains
5
Q
Order - mouse
A
Rodentia
6
Q
Clade - mouse
A
M. musculus
7
Q
Superfamily - Mouse
A
Muroidea
8
Q
Family - Mouse
A
Muridae
9
Q
House mouse clade
A
M. m. domesticus, M. m. musculus, M.m. castaneus, M. m. bactrianus, M.m. gentilulus
10
Q
Genetic mosaic of lab mouse
A
M.m. domesticus (60%), M.m. musculus (30%), and M.m. castaneus. Most lab mice share common mitochondrial genome from M.m. domesticus and common Y chromosome from M.m. musculus
11
Q
Abbie Lathrop’s Granby Mouse Farm, MA
A
Most lab mice strains derived from small number of fancy mice from a single breeder
12
Q
Mouse chromosome number - haploid
A
20, telocentric chromosomes
13
Q
MHC chr. location
A
17
14
Q
Coisogenic
A
Inbred strains that differ from original strain by mutation at single locus E.g. - 129S7/SvEvBrd- Fyntm1Sor Strain then DASH and gene
15
Q
Recombinent inbred strains
A
Sets of inbred mice derived from crossing two inbred strains, and then developed by single pair random mating of sibling mice from F2 generations. Each line maintained separately becoming its own inbred strain (20 generations) E.g. - CXB6, female first
16
Q
Outbred stock
A
E.g. - TAC:NIH Swiss COLON
17
Q
F1 Hybrid
A
E.g. - D2B6F1 Progeny of two inbred strains Uppercase abbreviations of parents followed by F1, genetically identical (F1 only)
18
Q
Congenic
A
One or more backcrosses to parental strain are made after F1 with selection for a particular marker (10 generations)
Recipient.donor-differential allele
E.g. - B6.AKR- H2 k
19
Q
Conplastic
A
Inbred mice that are congenic for mitochondrial DNA contributed by other strains. Nuclear genome from male of one strain backcrossed (at least 10) onto cytoplasmic genome of female of another
E.g. - C57BL/6-mtBALB/c
NUCLEAR GENOME-mtcytoplasmic genome
20
Q
Consomic
A
Inbred mice that are congenic for an entire chromosome. Repeat backcrossing of whole chromosome onto an inbred strain (10 backcrosses) E.g. - SHR-ChrYBN HOSTSTRAIN-Chr#DONOR STRAIN
21
Q
Point mutations
A
ENU, EMS
22
Q
Cre
A
Coliphage F1, site specific recombinase loxP (target site)
23
Q
FLP
A
Saccharomyces cerevisiae, site specific recombinase FRT (target site)
24
Q
RNAi
A
Gene knockdown mice Electroporation or lentiviral infection
25
Original C57BL/6 mouse
Originated from Abbie Lantrop's female 57 (and male 52) at Cold spring Harbor Laboratory (C) and was the black (BL) line of this female
26
Mouse food consumption
3-5g/day after weaning
27
Water consumption - mouse
6-7mL/day
28
Eyes open - mice
12-13 days
29
TNZ mouse
29.6-30.5C
30
Mouse lung lobes
4 (right): superior, middle, inferior, postcaval
31
Mouse adaptation for high BMR
High alveolar PO2, rapid respiratory rate, short air passage, moderately high RBC count, high Hb and CA; high blood O2 capacity; high capillary density, high glucose,; more pronounced Bohr effect
32
Protein in mouse urin
Taurine is always present, tyrptophan absent Creatinine is excreted (not like other mammals)
33
Submaxillary gland - mouse
Secretes only seromucoid (mixed gland in most other mammals)
34
Esophagus of the mouse
Thick cornified squamous epithelium
35
Gastric mucosa - mice
Proximal portion of the stomach is keratinized, distal part is glandular
36
Order, suborder, superfamily, family, subfamily, genus, species of lab mouse
Rodentia, myomorpha, muroidea, muridae, murinae, mus, musculus
37
Mouse chromosome pairs and characteristics
20 telocentric chromosomes
38
Histocompatibility (H) loci
control expression of cell surface molecules that modulate critical immune responses
39
MHC chromosome
17
40
Inbred
Strains, 20 or more brother x sister (filial, F) generations
Not FULLY inbred until 60 F
41
Substrain
If branches of an inbred strain are separated before F40, if branches have separated for 100 generations, or if genetic differences arise
Base strain/ILAR lab code
42
Isogenic
When two inbred mouse strains are bred to each other the F1 hybrids are genetically identical to each other, but maximally heterozygous (with chromosomes of each chromosomal pair separately contributed by each parental strain)
43
F2 hybrids
Maximally genetically diverse from one another (with chromosomes of both chromosomal pairs containing a mixture of contributions from each parental strain)
44
Recombinant Inbred (RI)
Sets of inbred strains of mice derived from crossing two inbred strains, developed by single-pair random matings of sibling mice from the F2 generation
Propegated by brother-sister matings for 20 generations, with each line becoming a separate inbred strain
Useful for mapping phenotypic or quantitative traits that differ between the progenitor strains
45
Recombinant congenic strains
Sets of inbred strains with one or more backcrosses to one parental strain (background strain) made after the F1 generation, before inbreeding has begun
BackgroundStraincDonorStrain1
eg - CcS1
46
Advanced intracross lines (AILs)
RI line
Produce F2 generation between two inbred strains and then, in each subsequent generation, intercrossing mice but avoiding sibling matings
Increase possibility of recombination between tightly linked genes
LabCode:InbredStrain,InbredStrain-G#
47
Co-isogenic
When a mutation arises spontaneously or is induced within an inbred strain the mutant mouse becomes co-isogenic with the parent inbred strain
Identical except for one mutant allele
Strain-*differential allele*
C3H/HeJ-*md2J*
48
Backcrossing, congenic
F1 hybrid created by mating the donor mutant strain to the desired background strain while retaining the mutant locus
After 10 backcross generations the mutant mouse line is now congenic
Never coisogenic
49
F1 hybrids
Mice from crosses between inbred strains
Cannot be perpetuated
50
Segregating inbred strain
Brother-sister matings system for more than 20 generations with heterozygosity for the mutations forced by 1) backcrossing, 2) intercrossing, 3) crossing and intercrossing or 4) backcrossing and intercrossing
Continue brother-sister matings with heterozygosity force by above methods or with homozygosity forced by intercrossing homozygotes
51
Consomic strains
Aka - chromsome substitution strains
Inbred mice that are congenic for entire chromosomes
Useful for studying polygenic traits
52
Conplastic strains
Inbred mice that are congenic for different mitochondrial genomes contributed by other inbred strains, species, or subspecies
53
Mutagenic chemical that induce point mutations
ethylmethanesulphonate (EMS) or N-ethyl-N-nitrosurea (ENU)
54
Transgenic founders
Hemizygous - injected DNA is not homologous to the mouse genome and is not an allele
Each embryo genertically altered by additive transgenesis (pronuclear injection)
55
Gene trapping
High-throughput approach that randomly introduce insertional mutations within the genome
Vectors containing a gene trapping cassette with a promotor-less reporter gene and/or selectable genetic marker flanked by an upstream 3' splice site and a downstream termination sequence
Simultaneously inactive and report the expression of the trapped gene at the insertion site, and provide a DNA tag for the rapid identification of the disrupted genes
56
Targeted gene mutations
Homologous recombination of specific sites within the genome of the ES cells
Homologous sequences flank the upstream and downstream regions of the targeted gene and the construct between the flanking sequences may inactivate (knock out) or replace (knock in) a gene
57
Cre
Recombinase
from coliphage P1
58
FLP
Recombinase
from Saccharomyces cerevisiae
Mediates recombination via FRT (target site)
59
ES cell transgenics
Transformed ES cells are injected in the inner cell mass of recipient blastocysts, which are implanted into the uteri of pseudopregnant surrogate mothers
Pups that are born are a mix of cells derived from recipient blastocysts and transformed ES cells (chimeras)
60
RNAi
dsRNA used to produce gene knockdown mice
Proccessed into siRNAs which find a homologous target mRNA resulting in interference
61
ZFNs and TALENs
Engineered proteins that target DNA fused to the nonspecific endonuclease Fok1
62
CRIPSR
Clustered regularly interspaced short palindromic repeats
Target specific DNA sequences
Cas proteins produce DSB (double-stranded breaks)
63
Mechanism of EE's (engineered endonucleases)
Produce DSB by either NHEJ or homologous recombination (HR)
Vectors encoding EEs can be injected by pronuclear injection of DNA, intracytoplasmic injection of RNA, or transfection of mouse ES cells
64
NHEJ
Error-prone repair system
Results in insertions or deletions (indels) with a high frequency, which can result in gene disruption
65
HR
Less common repair pathway
Certain manipulations can be used to increase HR efficiency
Allows for introduction of donor DNA to generate knock-ins, specific point mutations, or for the generation of larger modifications such as insertion of loxP sites
66
F1 hybrid nomenclature
StrainStrain
Female partner listed first
C57Bl/6J129S2/SvPasF1
B6129F1
67
RI nomenclature
Parent Strain X Parent Strain - specific line
C57BL/6JXDBA/2J-1
BXD-1
68
Nomenclature - mutant genes
Brief abbreviation for the mutation
Parent gene in italics
Mutant gene in superscript
E.g - *LystbgJ*
69
Transgenic strain - nomenclature
Tg(YYY)#Zzz
Tg = transgenic
YYY = brief description of the inserted DNA
= series of event generated using a given construct
Zzz = lab code
E.g. - FVB/N-Tg(MMTV-Erb2)1Led
Inbred FVB/N mice in which the rat Erb2 gene was inserted under control of the mammary tumor virus LTR promotor, the first line created in the laboratory of Phil Leder
70
Transgene insertional mutation - nomenclature
Mutant allele of gene is given designation by using the gene symbol and an abbreviation for the transgene as a superscript (-*Abc*tg1Zzz*)*
71
Targeted mutation/KO nomenclature
Designated by the mutated gene with the identification of the mutational event as a superscript
*Cftrtm1Unc*
Targeted mutation (tm), first line (1) created at the University of North Carolina (Unc) in the cystic fibrosis transmembrane regulator gene
72
Congenic nomenclature
Background inbred strain **.** donor strain
C57BL/6N.129P2/OlaHsd-*Abctm1Zzz*
If the background strain is mixed genetic origin: **STOCK**.donor
If the donor strain is mixed: Cg
73
Incipient congenic
Backcrossing is incomplete but at the N5 generation
background inbred strain ; donor strain
74
Mixed Inbred
Strain derived from two strains which is not yet inbred
Strain1;Strain2
75
Pronuclear injection
Randomly inserted DNA, 1-5% success, multiple copies often inserted (concatamer), must integrate stably into at least one chromosomes to be genetically transmitted
Male mice are mated with young, superovulated female mice. Females are euthanized 0.5 DPC and embryos harvested. Injected and implanted into oviducts of 0.5 DPC pseudopregnant recipient.
DNA of interest is cloned within a bacterial plasmid and propogated in E. coli
Start sequence, promotor, cloned gene, stop sequence, and vector backbone
76
Blastocyst injection
Targeted mutation, ESCs mediated homologous recombination, screen chimeras for germline transmission
77
ENU
N-ethyl-N-nitrosurea
Targets the SSC in males and induces one mutation every 175-4500 loci
100mg/kg q3 weeks
Fertility recovery 12-20 weeks
