Molecular characterization of H3.3 G34 oncohistone mutations:
Gespeichert in:
| Beteilige Person: | |
|---|---|
| Format: | Hochschulschrift/Dissertation Buch |
| Sprache: | Englisch |
| Veröffentlicht: |
Heidelberg
2023
|
| Schlagwörter: | |
| Links: | https://d-nb.info/1289691673/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034574274&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| Umfang: | xix, 142 Seiten Illustrationen, Diagramme 30 cm |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV049313192 | ||
| 003 | DE-604 | ||
| 007 | t| | ||
| 008 | 230905s2023 gw a||| m||| 00||| eng d | ||
| 015 | |a 23,H09 |2 dnb | ||
| 016 | 7 | |a 1289691673 |2 DE-101 | |
| 020 | |c Broschur | ||
| 035 | |a (OCoLC)1378163730 | ||
| 035 | |a (DE-599)DNB1289691673 | ||
| 040 | |a DE-604 |b ger |e rda | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c XA-DE | ||
| 049 | |a DE-355 | ||
| 084 | |8 1\p |a 616.994 |2 23ksdnb | ||
| 084 | |8 3\p |a 570 |2 23sdnb | ||
| 084 | |8 2\p |a 610 |2 23sdnb | ||
| 100 | 1 | |a Kühn, Alexander |d 1992- |e Verfasser |0 (DE-588)1289124221 |4 aut | |
| 245 | 1 | 0 | |a Molecular characterization of H3.3 G34 oncohistone mutations |c presented by Alexander Kühn, M. Sc. |
| 264 | 1 | |a Heidelberg |c 2023 | |
| 300 | |a xix, 142 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 Ruprecht-Karls-University Heidelberg |d 2023 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 710 | 2 | |a Universität Heidelberg |0 (DE-588)2024349-2 |4 dgg | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1289691673/04 |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=034574274&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 883 | 0 | |8 1\p |a emakn |c 0,05210 |d 20230525 |q DE-101 |u https://d-nb.info/provenance/plan#emakn | |
| 883 | 2 | |8 2\p |a dnb |d 20230809 |q DE-101 |u https://d-nb.info/provenance/plan#dnb | |
| 883 | 2 | |8 3\p |a dnb |d 20230809 |q DE-101 |u https://d-nb.info/provenance/plan#dnb | |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-034574274 | |
Datensatz im Suchindex
| _version_ | 1822127864221270016 |
|---|---|
| adam_text |
CONTENT
TABLE
OF
CONTENTS
CONTRIBUTIONS
.
I
SUMMARY
.
III
ZUSAMMENFASSUNG
.
V
CONTENT
.
VII
LIST
OF
FIGURES
.
XI
LIST
OF
TABLES
.
XIII
LIST
OF
SUPPLEMENTARY
TABLES
.
XIII
ABBREVIATIONS
.XV
1.
INTRODUCTION
.
1
1.1
EPIGENETICS
AND
CHROMATIN
.
1
1.2
HISTONES
.
3
1.3
POST-TRANSLATIONAL
MODIFICATIONS
OF
HISTONES
AND
THEIR
ROLE
IN
GENE
REGULATION
.
5
1.4
THE
HISTONE
H3
FAMILY
.
8
1.4.1
THE
NON-CANONICAL
HISTONE
H3.3
VARIANT
.
8
1.4.2
DEPOSITION
OF
H3
VARIANTS
INTO
CHROMATIN
BY
CHAPERONES
.
10
1.5
DNA
METHYLATION
.
13
1.6
EPIGENETICS
OF
STEM
CELLS
AND
THEIR
DIFFERENTIATION
.
15
1.7
DEREGULATION
OF
THE
EPIGENOME
IN
CANCER
.
17
1.7.1
HISTONE
MUTATIONS
OCCUR
FREQUENTLY
IN
CANCER
.
19
1.7.2
H3.3
G34
MUTATIONS
AND
THEIR
ROLE
IN
CANCER
.
21
2.
AIMS
OF
THIS
STUDY
.
24
3.
MATERIALS
AND
METHODS
.
26
3.1
MATERIALS
.
26
3.1.1
CHEMICALS
.
26
3.1.2
CONSUMABLES
.
28
3.1.3
KITS
.
29
3.1.4
EQUIPMENT
.
30
3.1.5
BUFFERS
.
31
3.1.6
ANTIBIOTICS
.
33
3.1.7
ENZYMES
.
33
3.1.8
CELL
LINES
.
34
VII
3.1.9
CELL
CULTURE
MEDIA
AND
ADDITIVES
.
34
3.1.10
PRIMERS
AND
OLIGONUCLEOTIDES
.
36
3.1.10.1
MASSARRAY
PRIMERS
.
36
3.1.10.2
PRIMER
SEQUENCES
FOR
REAL-TIME
QUANTITATIVE
POLYMERASE
CHAIN
REACTION
(RT
QPCR)
36
3.1.10.3
PRIMERS
USED
FOR
SITE-DIRECTED
MUTAGENESIS
(SDM)
.
37
3.1.10.4
PRIMERS
AND
OLIGONUCLEOTIDES
USED
FOR
ANTIBODY-GUIDED
CHROMATIN
TAGMENTATION
(ACT-SEQ)
.
38
3.1.11
ANTIBODIES
.
39
3.1.12
SOFTWARE
.
41
3.2
METHODS
.
42
3.2.1
CELL
CULTURE
.
42
3.2.1.1
CULTIVATION
OF
HEK293T
.
42
3.2.1.2
CULTIVATION
OF
KH2
MESC
.
42
3.2.1.2.1
GENERATION
OF
MOUSE
EMBRYONIC
FIBROBLAST
(MEF)
FEEDER
CELLS
.
42
3.2.1.2.2
MAINTENANCE
CULTURE
OF
KH2
MESC
.
43
3.2.1.2.3
PRE-COATING
OF
CULTURE
PLATES
.
44
3.2.1.2.4
SEPARATION
OF
KH2
MESCS
AND
MEF
FEEDER
CELLS
.
44
3.2.1.2.5
TARGETING
OF
KH2
MESCS
.
44
3.2.1.2.6
VERIFICATION
OF
SINGLE
COPY
INTEGRATION
IN
KH2
MESCS
.
45
3.2.1.3
EMBRYOID
BODY
FORMATION
.
45
3.2.1.3.1
EMBRYOID
BODY
FORMATION
FOR
MRNA
EXPRESSION
ANALYSIS
.
45
3.2.1.3.2
GENERATION
OF
EMBRYOID
BODIES
FOR
HISTONE
EXTRACTION
.
46
3.2.2
LENTIVIRAL
TRANSDUCTION
.
46
3.2.2.1
LENTIVIRUS
PRODUCTION
.
46
3.2.2.2
LENTIVIRAL
TRANSDUCTION
AND
PUROMYCIN-MEDIATED
SELECTION
OF
HEK293T
CELLS
.
47
3.2.3
CLONING
OF
PLASMIDS
.
47
3.2.3.1
GENERATION
OF
PLVX
PLASMIDS
ENCODING
H3.3
VARIANTS
BY
SITE-DIRECTED
MUTAGENESIS
.
47
3.2.3.2
GENERATION
OF
PCTGIL
TARGETING
PLASMIDS
.
48
3.2.4
PRECIPITATION
OF
DNA.
.
50
3.2.5
BACTERIAL
TRANSFORMATION
.
50
3.2.6
PREPARATION
OF
GLYCEROL
STOCKS
.
50
3.2.7
PLASMID
PREPARATION
FROM
GLYCEROL
STOCKS
.
51
3.2.8
AGAROSE
GEL
ELECTROPHORESIS
.
51
VIII
3.2.9
ISOLATION
OF
RNA
AND
DNA
.
52
3.2.9.1
COMBINED
ISOLATION
OF
RNA
AND
DNA
FROM
ONE
SAMPLE
.
52
3.2.10
QUANTIFICATION
OF
DNA
AND
RNA
AND
QUALITY
CONTROL
.
54
3.2.11
SYNTHESIS
OF
COMPLEMENTARY
DNA
(CDNA)
.
54
3.2.12 EXPRESSION
ANALYSIS
USING
REAL-TIME
QUANTITATIVE
POLYMERASE
CHAIN
REACTION
(RT
QPCR)
.
55
3.2.13
TARGETED
ANALYSIS
OF
DNA
METHYLATION
USING
MASSARRAY
.
55
3.2.14
HISTONE
EXTRACTION
.
56
3.2.15
PROTEIN
ISOLATION
.
57
3.2.16
SUBCELLULAR
FRACTIONATION
.
57
3.2.17
DETERMINATION
OF
PROTEIN
CONCENTRATION
BY
BICINCHONINIC
ACID
(BCA)
ASSAY
.
58
3.2.18
SEPARATION
OF
PROTEINS
USING
SODIUM
DODECYL
SULFATE-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
(SDS-PAGE)
.
58
3.2.19
WESTERN
BLOT
.59
3.2.20
TESTING
OF
ANTIBODIES
USING
PEPTIDE
ARRAYS
.
59
3.2.21
ANTIBODY-GUIDED
CHROMATIN
TAGMENTATION
FOLLOWED
BY
SEQUENCING
(ACT-SEQ)
.
61
3.2.21.1
ANTIBODY-GUIDED
CHROMATIN
TAGMENTATION
(ACT)
.
61
3.2.21.2
LIBRARY
PREPARATION
FOR
NEXT-GENERATION
SEQUENCING
.62
3.2.21.3
PURIFICATION
OF
DNA
LIBRARIES
FOR
NEXT-GENERATION
SEQUENCING
.63
3.2.21.4
ACT-SEQ
DATA
ANALYSIS
.
64
3.2.22
STATISTICAL
ANALYSIS
.
65
3.2.23
VISUALISATION
.
65
4.
RESULTS
.
66
4.1
HISTONE
H3
PTMS
ARE
AFFECTED
BY
H3.3
G34
MUTATIONS
IN
CIS
.66
4.2
GENERATION
OF
A
MOUSE
EMBRYONIC
STEM
CELL
MODEL
SYSTEM
.
72
4.3
EXPRESSION
OF
STEM
CELL
MARKER
GENES
IN
MESCS
IS
NOT
CHANGED
BY
H3.3
G34
MUTANTS
.78
4.4
GLOBAL
LEVELS
OF
H3
POST-TRANSLATIONAL
MODIFICATIONS
ARE
NOT
AFFECTED
BY
EXPRESSION
OF
H3.3
G34
MUTANTS
IN
MESCS
.
79
4.5
CHROMATIN
PROFILING
OF
MESCS
.
82
4.6
DNA
METHYLATION
OF
REPETITIVE
ELEMENTS
IS
NOT
AFFECTED
BY
H3.3
G34
MUTANTS
IN
MESCS
.90
4.7
EFFECTS
OF
H3.3
G34
MUTANTS
ON
DIFFERENTIATION
.
92
4.7.1
.
DIFFERENTIATION
OF
MESCS
TO
EMBRYOID
BODIES
IS
NOT
IMPAIRED
IN
H3.3
G34
MUTANT
MESCS
.
92
4.7.2
GLOBAL
LEVELS
OF
H3
POST-TRANSLATIONAL
MODIFICATIONS
ARE
NOT
ALTERED
UPON
EMBRYOID
BODY
FORMATION
IN
H3.3
G34
MUTANT-EXPRESSING
MESCS
.
96
5.
DISCUSSION
.
101
IX
5.1
H3K36
METHYLATION
IS
REDUCED
ON
H3.3
G34
ONCOHISTONES
IN
CIS
.
102
5.2
GENERATION
OF
MESC
TO
STUDY
H3.3
G34
MUTATIONS
.
105
5.3
H3.3
G34
ONCOHISTONES
DO
NOT
ALTER
GLOBAL
LEVELS
OF
H3
MODIFICATIONS
IN
MESCS
.
106
5.4
CHROMATIN
PROFILING
OF
H3.3
G34
ONCOHISTONE-EXPRESSING
MESCS
.
108
5.5
H3.3
G34
ONCOHISTONES
DID
NOT
AFFECT
DIFFERENTIATION
OF
MESCS
TO
EMBRYOID
BODIES
.
111
5.6
CONCLUSIONS
&
OUTLOOK
.
113
6.
REFERENCES
.
117
7.
APPENDIX
.
133
7.1
SUPPLEMENTARY
TABLES
.
133
8.
PUBLICATIONS
AND
POSTER
PRESENTATIONS
.
138
9.
ACKNOWLEDGEMENTS
140
X |
| any_adam_object | 1 |
| author | Kühn, Alexander 1992- |
| author_GND | (DE-588)1289124221 |
| author_facet | Kühn, Alexander 1992- |
| author_role | aut |
| author_sort | Kühn, Alexander 1992- |
| author_variant | a k ak |
| building | Verbundindex |
| bvnumber | BV049313192 |
| ctrlnum | (OCoLC)1378163730 (DE-599)DNB1289691673 |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>00000nam a2200000 c 4500</leader><controlfield tag="001">BV049313192</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="007">t|</controlfield><controlfield tag="008">230905s2023 gw a||| m||| 00||| eng d</controlfield><datafield tag="015" ind1=" " ind2=" "><subfield code="a">23,H09</subfield><subfield code="2">dnb</subfield></datafield><datafield tag="016" ind1="7" ind2=" "><subfield code="a">1289691673</subfield><subfield code="2">DE-101</subfield></datafield><datafield tag="020" ind1=" " ind2=" "><subfield code="c">Broschur</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)1378163730</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)DNB1289691673</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rda</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="044" ind1=" " ind2=" "><subfield code="a">gw</subfield><subfield code="c">XA-DE</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-355</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="8">1\p</subfield><subfield code="a">616.994</subfield><subfield code="2">23ksdnb</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="8">3\p</subfield><subfield code="a">570</subfield><subfield code="2">23sdnb</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="8">2\p</subfield><subfield code="a">610</subfield><subfield code="2">23sdnb</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Kühn, Alexander</subfield><subfield code="d">1992-</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)1289124221</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Molecular characterization of H3.3 G34 oncohistone mutations</subfield><subfield code="c">presented by Alexander Kühn, M. Sc.</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Heidelberg</subfield><subfield code="c">2023</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">xix, 142 Seiten</subfield><subfield code="b">Illustrationen, Diagramme</subfield><subfield code="c">30 cm</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="b">Dissertation</subfield><subfield code="c">Ruprecht-Karls-University Heidelberg</subfield><subfield code="d">2023</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="710" ind1="2" ind2=" "><subfield code="a">Universität Heidelberg</subfield><subfield code="0">(DE-588)2024349-2</subfield><subfield code="4">dgg</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">B:DE-101</subfield><subfield code="q">application/pdf</subfield><subfield code="u">https://d-nb.info/1289691673/04</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034574274&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="883" ind1="0" ind2=" "><subfield code="8">1\p</subfield><subfield code="a">emakn</subfield><subfield code="c">0,05210</subfield><subfield code="d">20230525</subfield><subfield code="q">DE-101</subfield><subfield code="u">https://d-nb.info/provenance/plan#emakn</subfield></datafield><datafield tag="883" ind1="2" ind2=" "><subfield code="8">2\p</subfield><subfield code="a">dnb</subfield><subfield code="d">20230809</subfield><subfield code="q">DE-101</subfield><subfield code="u">https://d-nb.info/provenance/plan#dnb</subfield></datafield><datafield tag="883" ind1="2" ind2=" "><subfield code="8">3\p</subfield><subfield code="a">dnb</subfield><subfield code="d">20230809</subfield><subfield code="q">DE-101</subfield><subfield code="u">https://d-nb.info/provenance/plan#dnb</subfield></datafield><datafield tag="943" ind1="1" ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-034574274</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV049313192 |
| illustrated | Illustrated |
| indexdate | 2025-01-24T11:02:23Z |
| institution | BVB |
| institution_GND | (DE-588)2024349-2 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-034574274 |
| oclc_num | 1378163730 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | xix, 142 Seiten Illustrationen, Diagramme 30 cm |
| publishDate | 2023 |
| publishDateSearch | 2023 |
| publishDateSort | 2023 |
| record_format | marc |
| spelling | Kühn, Alexander 1992- Verfasser (DE-588)1289124221 aut Molecular characterization of H3.3 G34 oncohistone mutations presented by Alexander Kühn, M. Sc. Heidelberg 2023 xix, 142 Seiten Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Ruprecht-Karls-University Heidelberg 2023 (DE-588)4113937-9 Hochschulschrift gnd-content Universität Heidelberg (DE-588)2024349-2 dgg B:DE-101 application/pdf https://d-nb.info/1289691673/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034574274&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p emakn 0,05210 20230525 DE-101 https://d-nb.info/provenance/plan#emakn 2\p dnb 20230809 DE-101 https://d-nb.info/provenance/plan#dnb 3\p dnb 20230809 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Kühn, Alexander 1992- Molecular characterization of H3.3 G34 oncohistone mutations |
| subject_GND | (DE-588)4113937-9 |
| title | Molecular characterization of H3.3 G34 oncohistone mutations |
| title_auth | Molecular characterization of H3.3 G34 oncohistone mutations |
| title_exact_search | Molecular characterization of H3.3 G34 oncohistone mutations |
| title_full | Molecular characterization of H3.3 G34 oncohistone mutations presented by Alexander Kühn, M. Sc. |
| title_fullStr | Molecular characterization of H3.3 G34 oncohistone mutations presented by Alexander Kühn, M. Sc. |
| title_full_unstemmed | Molecular characterization of H3.3 G34 oncohistone mutations presented by Alexander Kühn, M. Sc. |
| title_short | Molecular characterization of H3.3 G34 oncohistone mutations |
| title_sort | molecular characterization of h3 3 g34 oncohistone mutations |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1289691673/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034574274&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT kuhnalexander molecularcharacterizationofh33g34oncohistonemutations AT universitatheidelberg molecularcharacterizationofh33g34oncohistonemutations |