Antisense and ribozyme methodology:
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
London [u.a.]
Chapman and Hall
1997
|
| Series: | Laboratory companion
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Literaturangaben |
| Physical Description: | XIII, 79 S. graph. Darst. |
| ISBN: | 3826100794 |
Staff View
MARC
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| 245 | 1 | 0 | |a Antisense and ribozyme methodology |c Ian Gibson (ed.) |
| 264 | 1 | |a London [u.a.] |b Chapman and Hall |c 1997 | |
| 300 | |a XIII, 79 S. |b graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 0 | |a Laboratory companion | |
| 500 | |a Literaturangaben | ||
| 650 | 4 | |a Antisense nucleic acids |v Laboratory manuals | |
| 650 | 4 | |a Catalytic RNA |v Laboratory manuals | |
| 650 | 4 | |a DNA, Antisense |x analysis |v Laboratory Manuals | |
| 650 | 4 | |a DNA, Antisense |x chemical synthesis |v Laboratory Manuals | |
| 650 | 4 | |a Genetic Techniques |v Laboratory Manuals | |
| 650 | 4 | |a RNA, Catalytic |x analysis |v Laboratory Manuals | |
| 650 | 4 | |a RNA, Catalytic |x chemical synthesis |v Laboratory Manuals | |
| 650 | 0 | 7 | |a Antisense-Nucleinsäuren |0 (DE-588)4282992-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Ribozym |0 (DE-588)4356469-0 |2 gnd |9 rswk-swf |
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| 689 | 1 | 0 | |a Ribozym |0 (DE-588)4356469-0 |D s |
| 689 | 1 | |5 DE-604 | |
| 700 | 1 | |a Gibson, Ian |e Sonstige |4 oth | |
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Record in the Search Index
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| adam_text | Contents
CHAPTER 1. Antisense and Ribozyme Methodology 1
1.1 The Potential 1
1.2 Antisense Technology 4
1.2.1 Problems 4
1.2.2 Resistance to Nucleases 4
1.2.3 Entry into Cells 5
1.2.4 How Antisense Works 6
1.2.5 Success 6
1.3 Ribozymes 7
1.3.1 What Are They? 7
1.3.2 Problems 8
1.3.3 Stable Ribozymes 8
1.3.4 Designing Ribozymes 8
1.4 Ribozymes or Antisense DNAs? 9
1.5 The Choice Today!! 10
CHAPTER 2. Design and Synthesis
of Antisense DNA Molecules 13
2.1 Introduction 13
2.2 Synthesis of Methylphosphonodiester Phosphodiester
Chimeric Oligodeoxynucleotides 15
2.2.1 Materials and Chemicals 15
2.2.2 Solutions 15
2.2.3 Maximizing Product Purity 16
2.2.4 Deprotection of Chimeric Oligodeoxynucleotides 16
2.2.5 Failed Sequences 17
2.3 Primary Purification by Reversed Phase, Solid Phase
Extraction on C18 SEP PAK Cartridges 17
2.3.1 Equipment 17
2.3.2 Method 18
2.3.3 Purification of the Oligodeoxynucleotide 18
2.3.4 Further Purification 19
2.4 Analysis and Purification by HPLC 20
2.4.1 Analysis of Chimeric Oligodeoxynucleotides by HPLC ... 20
Contents J/f
2.4.2 Purification of Chimeric Oligodeoxynucleotides
by HPLC 21
2.4.3 Re Use of Columns 22
2.5 Synthesis of Chimeric Oligodeoxynucleotides
with Fluorescein Attached 23
2.6 Summary 25
CHAPTER 3. The Design and Synthesis
of Hammerhead Ribozymes 27
3.1 Introduction 27
3.2 The Design of Hammerhead Ribozymes 29
3.3 Improving the Reactions 29
3.3.1 Accessibility of the Target Substrate Binding 29
3.3.2 Finding the Target 30
3.3.3 Theoretical Considerations 30
3.3.4 Experimental Approaches 31
3.3.5 Kinetic Studies 31
3.4 Length of Arms 33
3.4.1 Choosing Antisense Arms of Hammerhead Ribozymes. . . 33
3.4.2 Arms of Different Lengths 33
3.5 Cleavage of the Target Motif 34
3.6 Synthesis of Ribozymes 35
3.6.1 Chemical Synthesis of Short Hammerhead Ribozymes ... 36
3.6.2 Enzymatic Transcription in Vitro 36
3.7 Endogenous Expression of Ribozyme Genes 37
CHAPTER 4. Delivery of Ribozymes and Antisense
DNA Molecules into Mammalian Cells 41
4.1 Introduction 41
4.2 Exogenous Application 41
4.2.1 Intracytoplasmic Delivery of Antisense Oligodeoxy¬
nucleotides by Reversible Plasma Membrane Permea
bilization with Streptolysin O 43
4.3 Microinjection 44
4.4 Other Methods Used in Nucleic Acid Transfection 45
4.5 Electroporation 45
4.5.1 Method 45
4.5.2 Transfection: Optimization of Conditions 46
4.5.3 Mechanism of Uptake Following Electroporation 46
4.5.4 Benefits and Drawbacks of Electrophoretic Mediated
Uptake 47
4.6 Diethylaminoethyl Dextran (DEAE Dextran)
and DNA Transfection 47
4.6.1 Methods for Transfection of Adherent Cells 47
4.6.2 Possible Alterations of the Above Protocol 48
I
4.6.3 Transfection of Cells Growing in Suspension 49
4.6.4 Transfection Optimization 50
4.6.5 Mechanism of DEAE Dextran Uptake and Intracellular
Distribution 51
4.6.6 Benefits and Drawbacks 51
4.7 Calcium Phosphate Transfection 51
4.7.1 Method 52
4.7.2 Possible Alterations to Above Method 53
4.7.3 Method Optimization 54
4.7.4 Calcium Phosphate Mediated Uptake and Intracellular
Distribution 55
4.7.5 Benefits and Drawbacks of Calcium
Phosphate Mediated Uptake 56
4.8 Cationic Lipids 57
4.8.1 Cationic Lipid Formulations 57
4.8.2 Methods 58
4.8.3 Lipofectin Mediated Transient Transfection
of Adherent Cells 59
4.8.4 Lipofectin Mediated Stable Transfection
of Adherent Cells 59
4.8.5 Lipofectin Mediated Transfection of Cells
in Suspension 60
4.8.6 Lipofectamine Mediated Transient or Stable
Transfection of Adherent Cells 60
4.8.7 Lipofectamine Mediated Transfection of Cells
in Suspension 62
4.8.8 Optimizing Transfection 62
4.8.9 Cationic Lipid Uptake and Intracellular Distribution 64
4.8.10 Benefits and Drawbacks of Cationic Lipids 65
4.8.11 Future Developments 66
4.9 Vector Mediated Delivery 67
4.10 Conclusions 67
CHAPTER 5. The Future 73
Appendix 11
Subject Index 79
Contents gjjg
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| id | DE-604.BV011312052 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T18:07:37Z |
| institution | BVB |
| isbn | 3826100794 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-007599088 |
| oclc_num | 36767811 |
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| owner | DE-355 DE-BY-UBR DE-11 |
| owner_facet | DE-355 DE-BY-UBR DE-11 |
| physical | XIII, 79 S. graph. Darst. |
| publishDate | 1997 |
| publishDateSearch | 1997 |
| publishDateSort | 1997 |
| publisher | Chapman and Hall |
| record_format | marc |
| series2 | Laboratory companion |
| spelling | Antisense and ribozyme methodology Ian Gibson (ed.) London [u.a.] Chapman and Hall 1997 XIII, 79 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier Laboratory companion Literaturangaben Antisense nucleic acids Laboratory manuals Catalytic RNA Laboratory manuals DNA, Antisense analysis Laboratory Manuals DNA, Antisense chemical synthesis Laboratory Manuals Genetic Techniques Laboratory Manuals RNA, Catalytic analysis Laboratory Manuals RNA, Catalytic chemical synthesis Laboratory Manuals Antisense-Nucleinsäuren (DE-588)4282992-6 gnd rswk-swf Ribozym (DE-588)4356469-0 gnd rswk-swf Antisense-Nucleinsäuren (DE-588)4282992-6 s DE-604 Ribozym (DE-588)4356469-0 s Gibson, Ian Sonstige oth HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007599088&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Antisense and ribozyme methodology Antisense nucleic acids Laboratory manuals Catalytic RNA Laboratory manuals DNA, Antisense analysis Laboratory Manuals DNA, Antisense chemical synthesis Laboratory Manuals Genetic Techniques Laboratory Manuals RNA, Catalytic analysis Laboratory Manuals RNA, Catalytic chemical synthesis Laboratory Manuals Antisense-Nucleinsäuren (DE-588)4282992-6 gnd Ribozym (DE-588)4356469-0 gnd |
| subject_GND | (DE-588)4282992-6 (DE-588)4356469-0 |
| title | Antisense and ribozyme methodology |
| title_auth | Antisense and ribozyme methodology |
| title_exact_search | Antisense and ribozyme methodology |
| title_full | Antisense and ribozyme methodology Ian Gibson (ed.) |
| title_fullStr | Antisense and ribozyme methodology Ian Gibson (ed.) |
| title_full_unstemmed | Antisense and ribozyme methodology Ian Gibson (ed.) |
| title_short | Antisense and ribozyme methodology |
| title_sort | antisense and ribozyme methodology |
| topic | Antisense nucleic acids Laboratory manuals Catalytic RNA Laboratory manuals DNA, Antisense analysis Laboratory Manuals DNA, Antisense chemical synthesis Laboratory Manuals Genetic Techniques Laboratory Manuals RNA, Catalytic analysis Laboratory Manuals RNA, Catalytic chemical synthesis Laboratory Manuals Antisense-Nucleinsäuren (DE-588)4282992-6 gnd Ribozym (DE-588)4356469-0 gnd |
| topic_facet | Antisense nucleic acids Laboratory manuals Catalytic RNA Laboratory manuals DNA, Antisense analysis Laboratory Manuals DNA, Antisense chemical synthesis Laboratory Manuals Genetic Techniques Laboratory Manuals RNA, Catalytic analysis Laboratory Manuals RNA, Catalytic chemical synthesis Laboratory Manuals Antisense-Nucleinsäuren Ribozym |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007599088&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT gibsonian antisenseandribozymemethodology |