The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms:
Gespeichert in:
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Freiburg im Breisgau
[2021]
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MARC
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| 016 | 7 | |a 1230576266 |2 DE-101 | |
| 020 | |c Broschur | ||
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| 100 | 1 | |a Staehle, Hans Felix |d 1992- |e Verfasser |0 (DE-588)1227969899 |4 aut | |
| 245 | 1 | 0 | |a The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms |c vorgelegt 2020 von Hans Felix Staehle |
| 264 | 1 | |a Freiburg im Breisgau |c [2021] | |
| 300 | |a 159 Seiten |b Illustrationen, Diagramme |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Albert-Ludwigs-Universität Freiburg im Breigau |d 2021 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1230576266/04 |3 Inhaltsverzeichnis |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032833133&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-032833133 | ||
| 883 | 0 | |8 1\p |a aepgnd |c 0,12524 |d 20210416 |q DE-101 |u https://d-nb.info/provenance/plan#aepgnd | |
Datensatz im Suchindex
| _version_ | 1804182715891712000 |
|---|---|
| adam_text | LIST
OF
CONTENTS
.................................
-
...............
-
........................................
1
1
INTRODUCTION
........................................................................................................
5
1.1
HEMATOPOIESIS
..............................................................................
-
-
5
1.2
MYELOPROLIFERATIVE
NEOPLASMS
(MPNS)
.....................
7
1.2.1
WHO
CLASSIFICATIONS
----------------------------------7
1.2.2
PH-POSITIVE
MPNS
........................................ .
-
-
-----------
8
1.2.3
PH-NEGATIVE
MPNS
-
---------------
..............
---------
8
1.3
THE
TRANSCRIPTION
FACTOR
NUCLEAR
FACTOR
ERYTHROID
2
(NFE2)
..........................
19
1.3.1
STRUCTURE
AND
FUNCTION
OF
NFE2
-
-
...............................................
19
1.3.2
THE
ROLE
OF
NFE2
IN
MPNS
...............................................................
20
1.3.3
SIGNALLING
PATHWAYS
OF
NFE2
............
-
................
20
1.4
EPIGENETICS
-
.......................................................
-
......................................
-22
1.4.1
DNA
METHYLATION
-
-
.....................................................................
22
1.4.2
HISTONE
MODIFICATIONS
-
-
............
-
................................................
24
1.4.3
THE
HISTONE
DEMETHYLASE
JMJD1C
..................................................
28
1.4.4
THE
HISTONE
DEMETHYLASE
JMJD2C
......................
-
........................
30
1.5
AIMS
AND
SIGNIFICANCE
................
-
...............
-
...............................................
-31
2
MATERIAL
AND
METHODS
..........................................................
-
..............
33
2.1
CLONING
TECHNIQUES
AND
PLASMID
PREPARATION
.
...............................................
33
2.1.1
PLASMIDS
.............................
-
...........................................................
33
2.1.2
RESTRICTION
DIGEST
-
...........................................................................
34
2.1.3
VECTOR
DEPHOSPHORYLATION
-----------------------
----------------------
-
37
2.1.4
GEL
EXTRACTION
OF
DIGESTED
PLASMID
DNA
..............
-
.........................
-37
2.1.5
LIGATION
............................................................................................
38
2.1.6
TRANSFORMATION
OF
COMPETENT
CELLS
...................................................
39
2.1.7
MINI-PREPARATION
OF
PLASMID
DNA
(CTAB
METHOD)
...........................
41
2.1.8
MAXI-PREPARATION
OF
PLASMID
DNA
(QIAGEN)
. ................................
42
2.1.9
FREEZING
OF
BACTERIA
-
-
..................
-
............................................
43
2.1.10
SEQUENCING
....................................
-
..............................................
43
2.2
CELL
CULTURE
TECHNIQUES
-
...................................................
-
................
45
2.2.1
HEK
293T
CELLS
.........
-
.........................
-
-
.......................................
45
2.2.2
CELL
NUMBER
DETERMINATION
VIA
NEUBAUER
CHAMBER
.........................
46
2.2.3
FREEZING
AND
THAWING
OF
CELLS
..........................................................
47
1
2.3
MOUSE
STRAINS
-
-
.......................................
48
2.3.1
TRANSGENIC
MICE
........................
-
.....................................
-
............
48
2.3.2
KNOCKOUT
MICE
.................................................................................
50
2.3.3
JAK2
V617F
KNOCK-IN
MICE
---------------------------
-
-
..............
52
2.4
MOUSE
SPECIFIC
APPLICATIONS
...........................................................................
54
2.4.1
GENOTYPING
-
..............
-
..........
-
-
................................................
54
2.4.2
NON-OPERATIVE
PROCEDURES
ON
MICE
...........................
-
....................
54
2.4.3
BLOOD
SAMPLES
.................................................................................
56
2.4.4
BONE
MARROW
SAMPLES
.....................................................................
59
2.4.5
DISSECTION
................................
60
2.5
FLOW
CYTOMETRY
...............................................................................
-
.............
62
2.5.1
GENERAL
INFORMATION
-
...........
62
2.5.2
SIX-COLOR
PANEL
-
...........................................................................
62
2.5.3
SEVEN-COLOR
PANEL
................................
65
2.6
FURTHER
APPLICATIONS
FOR
PROTEINS
...................................................................
-67
2.6.1
PROTEIN
CONSERVATION
................
-
-
...............................................
67
2.6.2
DETERMINATION
OF
PROTEIN
CONCENTRATIONS
(BRADFORD
ASSAY)
..............
69
2.6.3
SODIUM-DODECYL-SULFATE-POLYACRYLAMIDE-GEL-ELECTROPHORESIS
-
70
2.6.4
WESTERN
BLOTTING
.................
72
2.7
FURTHER
APPLICATIONS
FOR
NUCLEIC
ACIDS
.............................................................
76
2.7.1
ISOLATION
OF
GENOMIC
DNA
AND
RNA
................
-
..............................
76
2.7.2
REVERSE
TRANSCRIPTION
............
-
.........................................................
77
2.7.3
POLYMERASE
CHAIN
REACTION
(PCR)
..............................
78
2.7.4
REVERSE
TRANSCRIPTION
QUANTITATIVE
POLYMERASE
CHAIN
REACTION
-
-
83
2.7.5
AGAROSE
GEL
ELECTROPHORESIS
.........
...................................................
84
2.7.6
QUANTIFICATION
OF
NUCLEIC
ACIDS
............................
-
86
2.8
STATISTICS
-
............
-
.......................................
-
...............................................
86
3
RESULTS
............................
87
3.1
VALIDATION
OF
JMJDLC
AND
JMJD2C
KNOCKOUT
MOUSE
MODELS
. ..........................
87
3.1.1
CLONING
PROJECTS
................
-
............................................................
87
3.1.2
ANTIBODY
TESTING
........................
93
3.1.3
VALIDATION
OF
JMJDLC^
MICE
.......................................
-94
3.1.4
VALIDATION
OF
JMJD2D
I
I
MICE
....................
-
-
................................
94
3.2
THE
HEMATOLOGICAL
PHENOTYPE
OF
JMJD1C
D,D
AND
JMJD2C
SL I
MICE
......................
97
3.2.1
EXPERIMENTAL
SET-UP
.
.......................................................................
97
3.2.2
THE
IN
VIVO
EFFECT
OF
JMJDLC
KNOCKOUT
ON
HEALTHY
HEMATOPOIESIS
-
-
97
3.2.3
THE
IN
VIVO
EFFECT
OF
JMJD2C
KNOCKOUT
ON
HEALTHY
HEMATOPOIESIS
-
-
97
2
3.3
KNOCKOUT
OF
JMJDLC
IN
JAK2
V617F
-DRIVEN
HEMATOPOIESIS
-
..............................
100
3.3.1
BREEDING
OF
JAK2
V617F
MX1-CRE
JMJD1C
D/D
MICE
..................................
100
3.3.2
THE
EFFECT
OF
JMJD1C
D/D
KNOCKOUT
IN
JA/C2
V6
7F
-DRIVEN
HEMATOPOIESIS
100
4
DISCUSSION
............................................................................................................
103
4.1
BREEDING
OF
JMJDLC
AND
JMJD2C
KNOCKOUT
MOUSE
LINES
..................................
103
4.1.1
JMJD1C
D/D
MICE
ARE
REAL
KNOCKOUTS
-------------------------------------------
103
4.1.2 JMJD2C
D/D
MICE
ARE
REAL
KNOCKOUTS
.
..................................................
104
4.2
THE
ROLE
OF
JMJDLC
AND
JMJD2C
IN
HEALTHY
HEMATOPOIESIS
...............................
104
4.2.1
KNOCKOUT
OF
JMJDLC
IS
DISPENSABLE
FOR
HEALTHY
HEMATOPOIESIS
-
-
104
4.2.2
KNOCKOUT
OF
JMJD2C
IS
DISPENSABLE
FOR
HEALTHY
HEMATOPOIESIS
-
-
105
4.3
THE
ROLE
OF
JMJDLC
AND
JMJD2C
IN
THE
CONTEXT
OF
MPNS
.................................
105
4.3.1
JMJDLC
IS
DISPENSABLE
FOR
JAK2
V617F
-DRIVEN
HEMATOPOIESIS
............
105
4.3.2
PREDICTIONS
ABOUT
THE
KNOCKOUT
OF
JMJD2C
IN
MPNS
-
.......................
107
4.4
CONCLUSION
....................................................................................................
-108
4.5
OUTLOOK
-
-
......................................................
108
5
SUMMARY-
.............................
-
.................
-
.........................................................
109
5.1
ENGLISH
................
-
.....................................
-
.................
-
....................
---
109
5.2
GERMAN
...........................................................................................................
110
LIST
OF
FIGURES
-
...............
-
-
.............................
-
-
-
.................................................
111
LIST
OF
TABLES
...............................
-
..........................................................................
113
ACRONYMS
...........
-
........................................................................
-
................
115
SUPPLEMENTAL
MATERIAL
.............................
-
.........................................................
121
REFERENCES
..................
-
.................
-
....................
123
ACKNOWLEDGEMENTS
..............................................................
-
.................
-
............
151
APPENDIX
................
-
...................
-
..........................................................
-
..............
-
153
3
|
| adam_txt |
LIST
OF
CONTENTS
.
-
.
-
.
1
1
INTRODUCTION
.
5
1.1
HEMATOPOIESIS
.
-
-
5
1.2
MYELOPROLIFERATIVE
NEOPLASMS
(MPNS)
.
7
1.2.1
WHO
CLASSIFICATIONS
----------------------------------7
1.2.2
PH-POSITIVE
MPNS
. .
-
-
-----------
8
1.2.3
PH-NEGATIVE
MPNS
-
---------------
.
---------
8
1.3
THE
TRANSCRIPTION
FACTOR
NUCLEAR
FACTOR
ERYTHROID
2
(NFE2)
.
19
1.3.1
STRUCTURE
AND
FUNCTION
OF
NFE2
-
-
.
19
1.3.2
THE
ROLE
OF
NFE2
IN
MPNS
.
20
1.3.3
SIGNALLING
PATHWAYS
OF
NFE2
.
-
.
20
1.4
EPIGENETICS
-
.
-
.
-22
1.4.1
DNA
METHYLATION
-
-
.
22
1.4.2
HISTONE
MODIFICATIONS
-
-
.
-
.
24
1.4.3
THE
HISTONE
DEMETHYLASE
JMJD1C
.
28
1.4.4
THE
HISTONE
DEMETHYLASE
JMJD2C
.
-
.
30
1.5
AIMS
AND
SIGNIFICANCE
.
-
.
-
.
-31
2
MATERIAL
AND
METHODS
.
-
.
33
2.1
CLONING
TECHNIQUES
AND
PLASMID
PREPARATION
.
.
33
2.1.1
PLASMIDS
.
-
.
33
2.1.2
RESTRICTION
DIGEST
-
.
34
2.1.3
VECTOR
DEPHOSPHORYLATION
-----------------------
----------------------
-
37
2.1.4
GEL
EXTRACTION
OF
DIGESTED
PLASMID
DNA
.
-
.
-37
2.1.5
LIGATION
.
38
2.1.6
TRANSFORMATION
OF
COMPETENT
CELLS
.
39
2.1.7
MINI-PREPARATION
OF
PLASMID
DNA
(CTAB
METHOD)
.
41
2.1.8
MAXI-PREPARATION
OF
PLASMID
DNA
(QIAGEN)
. .
42
2.1.9
FREEZING
OF
BACTERIA
-
-
.
-
.
43
2.1.10
SEQUENCING
.
-
.
43
2.2
CELL
CULTURE
TECHNIQUES
-
.
-
.
45
2.2.1
HEK
293T
CELLS
.
-
.
-
-
.
45
2.2.2
CELL
NUMBER
DETERMINATION
VIA
NEUBAUER
CHAMBER
.
46
2.2.3
FREEZING
AND
THAWING
OF
CELLS
.
47
1
2.3
MOUSE
STRAINS
-
-
.
48
2.3.1
TRANSGENIC
MICE
.
-
.
-
.
48
2.3.2
KNOCKOUT
MICE
.
50
2.3.3
JAK2
V617F
KNOCK-IN
MICE
---------------------------
-
-
.
52
2.4
MOUSE
SPECIFIC
APPLICATIONS
.
54
2.4.1
GENOTYPING
-
.
-
.
-
-
.
54
2.4.2
NON-OPERATIVE
PROCEDURES
ON
MICE
.
-
.
54
2.4.3
BLOOD
SAMPLES
.
56
2.4.4
BONE
MARROW
SAMPLES
.
59
2.4.5
DISSECTION
.
60
2.5
FLOW
CYTOMETRY
.
-
.
62
2.5.1
GENERAL
INFORMATION
-
.
62
2.5.2
SIX-COLOR
PANEL
-
.
62
2.5.3
SEVEN-COLOR
PANEL
.
65
2.6
FURTHER
APPLICATIONS
FOR
PROTEINS
.
-67
2.6.1
PROTEIN
CONSERVATION
.
-
-
.
67
2.6.2
DETERMINATION
OF
PROTEIN
CONCENTRATIONS
(BRADFORD
ASSAY)
.
69
2.6.3
SODIUM-DODECYL-SULFATE-POLYACRYLAMIDE-GEL-ELECTROPHORESIS
-
70
2.6.4
WESTERN
BLOTTING
.
72
2.7
FURTHER
APPLICATIONS
FOR
NUCLEIC
ACIDS
.
76
2.7.1
ISOLATION
OF
GENOMIC
DNA
AND
RNA
.
-
.
76
2.7.2
REVERSE
TRANSCRIPTION
.
-
.
77
2.7.3
POLYMERASE
CHAIN
REACTION
(PCR)
.
78
2.7.4
REVERSE
TRANSCRIPTION
QUANTITATIVE
POLYMERASE
CHAIN
REACTION
-
-
83
2.7.5
AGAROSE
GEL
ELECTROPHORESIS
.
.
84
2.7.6
QUANTIFICATION
OF
NUCLEIC
ACIDS
.
-
86
2.8
STATISTICS
-
.
-
.
-
.
86
3
RESULTS
.
87
3.1
VALIDATION
OF
JMJDLC
AND
JMJD2C
KNOCKOUT
MOUSE
MODELS
. .
87
3.1.1
CLONING
PROJECTS
.
-
.
87
3.1.2
ANTIBODY
TESTING
.
93
3.1.3
VALIDATION
OF
JMJDLC^
MICE
.
-94
3.1.4
VALIDATION
OF
JMJD2D
I
"
I
MICE
.
-
-
.
94
3.2
THE
HEMATOLOGICAL
PHENOTYPE
OF
JMJD1C
D,D
AND
JMJD2C
SL I
MICE
.
97
3.2.1
EXPERIMENTAL
SET-UP
.
.
97
3.2.2
THE
IN
VIVO
EFFECT
OF
JMJDLC
KNOCKOUT
ON
HEALTHY
HEMATOPOIESIS
-
-
97
3.2.3
THE
IN
VIVO
EFFECT
OF
JMJD2C
KNOCKOUT
ON
HEALTHY
HEMATOPOIESIS
-
-
97
2
3.3
KNOCKOUT
OF
JMJDLC
IN
JAK2
V617F
-DRIVEN
HEMATOPOIESIS
-
.
100
3.3.1
BREEDING
OF
JAK2
V617F
MX1-CRE
JMJD1C
D/D
MICE
.
100
3.3.2
THE
EFFECT
OF
JMJD1C
D/D
KNOCKOUT
IN
JA/C2
V6
'
7F
-DRIVEN
HEMATOPOIESIS
100
4
DISCUSSION
.
103
4.1
BREEDING
OF
JMJDLC
AND
JMJD2C
KNOCKOUT
MOUSE
LINES
.
103
4.1.1
JMJD1C
D/D
MICE
ARE
REAL
KNOCKOUTS
-------------------------------------------
103
4.1.2 JMJD2C
D/D
MICE
ARE
REAL
KNOCKOUTS
.
.
104
4.2
THE
ROLE
OF
JMJDLC
AND
JMJD2C
IN
HEALTHY
HEMATOPOIESIS
.
104
4.2.1
KNOCKOUT
OF
JMJDLC
IS
DISPENSABLE
FOR
HEALTHY
HEMATOPOIESIS
-
-
104
4.2.2
KNOCKOUT
OF
JMJD2C
IS
DISPENSABLE
FOR
HEALTHY
HEMATOPOIESIS
-
-
105
4.3
THE
ROLE
OF
JMJDLC
AND
JMJD2C
IN
THE
CONTEXT
OF
MPNS
.
105
4.3.1
JMJDLC
IS
DISPENSABLE
FOR
JAK2
V617F
-DRIVEN
HEMATOPOIESIS
.
105
4.3.2
PREDICTIONS
ABOUT
THE
KNOCKOUT
OF
JMJD2C
IN
MPNS
-
.
107
4.4
CONCLUSION
.
-108
4.5
OUTLOOK
-
-
.
108
5
SUMMARY-
.
-
.
-
.
109
5.1
ENGLISH
.
-
.
-
.
-
.
---
109
5.2
GERMAN
.
110
LIST
OF
FIGURES
-
.
-
-
.
-
-
-
.
111
LIST
OF
TABLES
.
-
.
113
ACRONYMS
.
-
.
-
.
115
SUPPLEMENTAL
MATERIAL
.
-
.
121
REFERENCES
.
-
.
-
.
123
ACKNOWLEDGEMENTS
.
-
.
-
.
151
APPENDIX
.
-
.
-
.
-
.
-
153
3 |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Staehle, Hans Felix 1992- |
| author_GND | (DE-588)1227969899 |
| author_facet | Staehle, Hans Felix 1992- |
| author_role | aut |
| author_sort | Staehle, Hans Felix 1992- |
| author_variant | h f s hf hfs |
| building | Verbundindex |
| bvnumber | BV047430652 |
| ctrlnum | (OCoLC)1238102862 (DE-599)KXP1749198878 |
| discipline | Medizin |
| discipline_str_mv | Medizin |
| format | Thesis Book |
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| id | DE-604.BV047430652 |
| illustrated | Illustrated |
| index_date | 2024-07-03T17:58:21Z |
| indexdate | 2024-07-10T09:11:58Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-032833133 |
| oclc_num | 1238102862 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | 159 Seiten Illustrationen, Diagramme 30 cm |
| publishDate | 2021 |
| publishDateSearch | 2021 |
| publishDateSort | 2021 |
| record_format | marc |
| spelling | Staehle, Hans Felix 1992- Verfasser (DE-588)1227969899 aut The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms vorgelegt 2020 von Hans Felix Staehle Freiburg im Breisgau [2021] 159 Seiten Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Albert-Ludwigs-Universität Freiburg im Breigau 2021 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1230576266/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032833133&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p aepgnd 0,12524 20210416 DE-101 https://d-nb.info/provenance/plan#aepgnd |
| spellingShingle | Staehle, Hans Felix 1992- The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms |
| subject_GND | (DE-588)4113937-9 |
| title | The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms |
| title_auth | The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms |
| title_exact_search | The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms |
| title_exact_search_txtP | The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms |
| title_full | The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms vorgelegt 2020 von Hans Felix Staehle |
| title_fullStr | The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms vorgelegt 2020 von Hans Felix Staehle |
| title_full_unstemmed | The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms vorgelegt 2020 von Hans Felix Staehle |
| title_short | The role of the histone demethylases Jmjd1c and Jmjd2c in the pathophysiology of myeloproliferative neoplasms |
| title_sort | the role of the histone demethylases jmjd1c and jmjd2c in the pathophysiology of myeloproliferative neoplasms |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1230576266/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032833133&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT staehlehansfelix theroleofthehistonedemethylasesjmjd1candjmjd2cinthepathophysiologyofmyeloproliferativeneoplasms |
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