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» Als Illustrator und Arzt übersetze ich Wissenschaft in Bilder.«
Dr. Guido Hegasy
www.hegasy.deWissenschaftliche IllustrationenFacharzt für Mikrobiologie
1 Bilharziose2 Multiple Sklerose3 Händehygiene4 Hautschichten5 Herzzyklus6 CRISPR-Cas97 Komplement 8 EHEC in Hamburg9 Immunantwort
1 2 3
5 6
7 8
4
9
Lebenszyklus eines Pärchen-Egels
(Schistosoma mansoni)
Freisetzung von Zerkarien
Schlüpfen der Mirazidien
Infektion von Schnecken
Freisetzung der Eiermit Stuhl und Urin
AdulteSchistosoma
Ei-Passage inDarm und Blase
Paarung
Produktionvon Eiern
Entwicklung vonSchistosomulae
Durchdringung der Haut
Illustrationhegasy.de
Doppelt-sehen
EinseitigerVisusverlust
Symptome und Befunde bei Multipler Sklerose
Intentionstremor Internucleäre Ophthalmoplegie
Periventrikuläre Entmarkungsherde
Lähmung,Schwäche,
Gehbehinderung
SpasmenBabinski ReflexInkontinenz
Verschwommen-sehen
3×
30 s
Hygienische Händedesinfektion
A
BC
D
EF
G
H
Leistenhaut (z.B. Handfläche)
1
2
3
4
5
7
12
1
3
4 7
9
12
6 8
10
11
Felderhaut (z.B. Achsel)
AC
D
EF
G
H
Schichten und Rezeptoren der menschlichen Haut
B – Stratum lucidum
A – Stratum corneum
C – Stratum granulosum
E – Stratum basale
D – Stratum spinosum
F – Stratum papillare
G – Stratum reticulare
H – Subcutis
Cutis Ep
ider
mis
Derm
is
10 Haarfollikel
M. arrector pili11
Fettgewebe12
Merkelzelle3
Meissner Körperchen2
Ruffini Körperchen4
Freie Nervenendigung1
Pacini Körperchen5
Haarfollikel-Sensor6
Holokrine Drüse9
Ekkrine Drüse7
Apokrine Drüse8
Arterie
Vene
Nerv
Konzept & Illustration:
hegasy.de
Illustrationhegasy.de
Der Herzzyklus
Vena cava inferior
Vena cava superior
Truncus pulmonalis
Aorta
Valva bicuspidalis
Valva aortae
Valva trunci pulmonalisChordaetendineae
Venae pulmonales
Arteriae pulmonales
Atrium dextrum
Atrium sinistrum
Musculuspapillaris
Truncus brachiocephalicus
Arteria carotis communis
Arteriasubclavia
Ventriculus dexter
Valvatricuspidalis Ventriculus
sinister
A.p.
V.p.
1
3
24
Mitral- & Trikuspidalklappe
Pulmonal- &Aortenklappe
Ventrikuläre Systole
Ventrikuläre Diastole
Ventil-Ebene
Ventrikulär Ejektion geschlossen offen2
Isovolumische Kontraktion geschlossen geschlossen1
Isovolumische Relaxation geschlossen geschlossen3
Ventrikuläre Füllung offen geschlossen4
EKG
Syst
ole
Dias
tole
Origin and Function of
CRISPR-Cas9 Technology
Introduction to CRISPR Biology:
Cas9 IntroducesTargeted DNA Breaks Panel
– 4 –
Origin and Function of
CRISPR-Cas9 Technology
Panel– 5 –
Origin and Function of
CRISPR-Cas9 Technology
Panel– 6 –
Exploiting Endogenous DNA Repair Mechanisms
Introduction to CRISPR Biology:
CRISPR-Cas9 Technology Allows Gene Engineering
Introduction to CRISPR Biology:
Double-Strand DNA BreaksAllow Gene Editing
NHEJInsertion Deletion
HDRTemplate Integration
Gene Disruption Gene Modification
Gene Editing
Fields of Applicationfor Cas9 Technology
Plan
ts Animals
H u m a ns
By joining tracrRNA and crRNA derived sequences with a linker loop, a simplified programmable tool for DNA manipulation is created. The tracrRNA-crRNA-chimera is called a single guide RNA (sgRNA). sgRNA can be easily synthesized. The complex, consisting of Cas9 and sgRNA, scans DNA for the presence of a protospacer adjacent motif (PAM). For Cas9 from Streptococcus pyogenes, this is a 5′-NGG-3′ sequence. When a PAM sequence is detected, the complementary DNA strand is compared to the crRNA derived guide region. If these sequences match, the DNA double strand is cleaved ~3 bp away of the PAM. This introduces a double-strand break (DSB). With both domains located in the NUC lobe of Cas9, the HNH domain cuts the strand complementary to the guide sequence (target strand) while the RuvC domain cuts the opposite strand. A DSB introduced precisely at a desired genetic site provides a tool for targeted genetic manipulations. References: Nishimasu H et al. CELL 2014, Hsu P et al. CELL 2014
When a double-strand break (DSB) is introduced in a cell’s DNA, there are two major repair pathways. In one pathway the blunt DNA ends are joined in a process called non-homologous end joining (NHEJ). This mechanism is error-prone and may produce insertions or deletions of bases. These alterations are collectively called Indel mutations. Indel mutations may result in a loss of function of the affected gene, e.g., by introducing a frameshift and/ or a premature stop codon.Another pathway is called homology directed repair (HDR). This pathway uses a DNA template to repair the site where the DSB occurred. When exogenous DNA is added, the sequence may serve as a template and is then integrated into the repair process. This may lead to the introduction of a new genetic sequence. Both repair pathways may result in a genetic alteration. Thus, targeted introduction of DSBs opens the door for gene editing. Reference: Doudna JA and Charpentier E SCIENCE 2014.
By introducing double-strand breaks into a living cell’s genomic DNA, CRISPR-Cas9 technology provides a potential molecular tool. This technology enables precise targeting of genes in live organisms to render them accessible to modifications. This technique has already been shown to work successfully in a variety of species and seems to be applicable to virtually any organism. In maize plants, CRISPR-Cas9 technology has been employed to modify genes to increase herbicide resistance. In mice, a multitude of mutants have been produced, often serving as models to analyze human diseases. Genetic modifications in animals are feasible in humans too. Data have already been published from experiments on human embryos. For this reason, a group of scientists, led by those who have invented the technique, have called for a public discussion on the responsible use of CRISPR-Cas9 technology.Reference: Baltimore D et al. SCIENCE 2015.
Cas9 as a Molecular Tool
Dr. med. HegasyLife Science Illustration
Dr. med. HegasyLife Science Illustration
Dr. med. HegasyLife Science Illustration
PAMRuvC
HNH
sgRNA
LinkerLoop
NUC Lobe
REC Lobe
C3
C2a
C4b C3b
C1qC1-Komplex
MASP
C4a
CP-/ LP-Konvertase
C4b2a3b
AP-Konvertase
C3bBb3b
C3(H2O) C3(H2O)B+FD
C3(H2O)Bb+P
FB FDBa
C2
C4
C2a
C4b
C9C9
C9
FD
+H2O
C2bC4a
KlassischerWeg
LektinWeg
AlternativerWeg
Das Komplement System
TerminalerWeg
Membran-Angriffs-Komplex
Amplifikations- schleife
MBL, Ficolin
C7C6
C2b
C8γ
β α
C5a
P
Bb
C3aC3
C9, C9, C9, …
C3a
C1rC1s
FD
FB
FDBa
C5
C5b
Ziel-Oberfläche
C3a
C3Bb
C3b C3b
C3
Bb
C3b C3b
C9
C9
C9
C5b
C7 C8C6
Properdin
Antikörper
MikrobielleZucker
Serine Protease
Thioester
C3aC4aC5a
C3b
C5aC3a
Anaphylatoxine
Chemotaxis
Opsonin
Illustrationhegasy.de
EHEC Outbreak Incidencesin Germany
EHEC Outbreak Incidences in Hamburg
0
10
20
30
40
50
60
70
20 21 22 23 24 25 26 27 28 29
Num
ber o
f Cas
es
Calendar Week 2011
Day of Outbreak Detection
EHEC: Onset of Symptoms
HUS: Onset of Symptoms
EHEC+HUS: Date of Notification
Epicurve
Date
Incidence Distribution
1125Districts
2219
19
114
2.1 – 4.0 Cases / 100000
0.0
4.1 – 6.0
6.1 – 12.0
0.1 – 2.0
>12.0
1.0 – 2.02.1 – 4.04.1 – 12.0>12.0
0.00.1 – 2.02.1 – 4.04.1 – 6.06.1 – 12.0>12.0
City of Hamburg
Late
ncy
[day
s]
0
10
20
30
40
16 - 22 May
23 - 29 May
30 May - 5 June
6 - 12 June
13 - 19 June
20 - 26 June
27 June - 3 July
Q3+1.5 IQR
Median
Q1-1.5 IQR
Q1
Q3
Outliers
Notification Latency
Fenugreek Sprouts
In 2011 an outbreak of enterohemorrhagic Escherichia coli occurred in Germany. The outbreak was largely caused by contaminated fenugreek sprouts. The northern part of Germany was greatly affected by the outbreak, and the City State of Hamburg had the highest incidences. Incidences ranged from zero for 22 districts to >12 for four districts. Notification latency increased when the peak of the outbreak had passed. The outbreak was detected when 98 persons had already fallen ill from the infection whereas public health authorities had been notified in only 4 cases. Reference: Tahden M et al., PLoS ONE 2016
EHEC O104:H4 Outbreak in Hamburg, Germany, 2011
Illustrationhegasy.de
Bacterium
A bacterium may enter the body through injured skin or by penetrating the endothelia of the respiratory or gastrointestinal tract. This may trigger an immune response.
A bacterium may enter the body through injured skin or by penetrating the endothelia of the respiratory or gastrointestinal tract. This may trigger an immune response.
When crossing endothelial barriers and entering human tissue an antigen-presenting cell (APC), e.g., a dendritic cell (DC), may detect the invader and endocytose bacterial antigens.
When crossing endothelial barriers and entering human tissue an antigen-presenting cell (APC), e.g., a dendritic cell (DC), may detect the invader and endocytose bacterial antigens.
Ii
MHC II
Lysosomes
Endosome
Peptide
Antigen-LoadedMHC II
Endosomes containing bacterial proteins are fused with lysosomes and antigenic peptides are released. These are then loaded on MHC class II molecules to present them on the cell surface.
Endosomes containing bacterial proteins are fused with lysosomes and antigenic peptides are released. These are then loaded on MHC class II molecules to present them on the cell surface.
Mature DCs present antigenic peptides on MHC II to the T cell receptor of naïve CD4 T cells together with co-stimulatory surface molecules. This activates the T cell to proliferate and differentiate.
Mature DCs present antigenic peptides on MHC II to the T cell receptor of naïve CD4 T cells together with co-stimulatory surface molecules. This activates the T cell to proliferate and differentiate.
MHC II TCR
CD3
CD28
CD80
T CellDC
T cells of the Th1 subtype activate macrophages to phagocytose and digest bacteria. B cells are stimulated to synthesize IgG for opsonization. Neutrophils are activated to act at the site of infection.
T cells of the Th1 subtype activate macrophages to phagocytose and digest bacteria. B cells are stimulated to synthesize IgG for opsonization. Neutrophils are activated to act at the site of infection.
Th1 Cell
Macrophage
B Cell
Neutrophil
IFNγ TNFα
CD40L
The DC migrates through lymphatic vessels to a nearby lymph node. Here, it presents bacterial antigens to T cells that have reached the same lymph node via blood vessels.
The DC migrates through lymphatic vessels to a nearby lymph node. Here, it presents bacterial antigens to T cells that have reached the same lymph node via blood vessels.
CD4
APC
DC T Cell
Lymph Node
1
2
3
4 5 6
A Simplified View on the Generation of a Th1 Immune Response
Wissenschaftliche Illustrationen Facharzt für Mikrobiologie
Rantzaustraße 33 22041 Hamburg
040 - 348 618 67 [email protected]
Dr. Guido Hegasy
www.hegasy.de