CN102702332B - 一对转录激活子样效应因子核酸酶l1和r2及其编码基因与应用 - Google Patents
一对转录激活子样效应因子核酸酶l1和r2及其编码基因与应用 Download PDFInfo
- Publication number
- CN102702332B CN102702332B CN201210163377.XA CN201210163377A CN102702332B CN 102702332 B CN102702332 B CN 102702332B CN 201210163377 A CN201210163377 A CN 201210163377A CN 102702332 B CN102702332 B CN 102702332B
- Authority
- CN
- China
- Prior art keywords
- seq
- pair
- sequence
- many nucleotide
- rpe65
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 34
- 101710163270 Nuclease Proteins 0.000 title abstract description 17
- 230000000694 effects Effects 0.000 title description 3
- 238000013518 transcription Methods 0.000 title description 2
- 230000035897 transcription Effects 0.000 title description 2
- 239000012190 activator Substances 0.000 title 1
- 101150116978 RPE65 gene Proteins 0.000 claims abstract description 18
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 15
- 230000008685 targeting Effects 0.000 claims abstract description 12
- 230000004048 modification Effects 0.000 claims abstract description 11
- 238000012986 modification Methods 0.000 claims abstract description 11
- 239000002773 nucleotide Substances 0.000 claims description 35
- 125000003729 nucleotide group Chemical group 0.000 claims description 35
- 230000004927 fusion Effects 0.000 claims description 16
- 108091008146 restriction endonucleases Proteins 0.000 claims description 15
- 239000013612 plasmid Substances 0.000 claims description 14
- 235000018102 proteins Nutrition 0.000 claims description 14
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 108091033319 polynucleotide Proteins 0.000 claims description 8
- 102000040430 polynucleotide Human genes 0.000 claims description 8
- 239000002157 polynucleotide Substances 0.000 claims description 8
- 238000010363 gene targeting Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 230000003321 amplification Effects 0.000 claims description 2
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 abstract description 27
- 108090000790 Enzymes Proteins 0.000 abstract description 26
- 108020004414 DNA Proteins 0.000 abstract description 25
- 238000010459 TALEN Methods 0.000 abstract description 24
- 108010043645 Transcription Activator-Like Effector Nucleases Proteins 0.000 abstract description 24
- 102000053602 DNA Human genes 0.000 abstract description 4
- 101100528986 Homo sapiens RPE65 gene Proteins 0.000 abstract 2
- 230000035772 mutation Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 27
- 229940088598 enzyme Drugs 0.000 description 26
- 239000012634 fragment Substances 0.000 description 18
- 239000012636 effector Substances 0.000 description 16
- 230000029279 positive regulation of transcription, DNA-dependent Effects 0.000 description 15
- 238000010586 diagram Methods 0.000 description 12
- 108091028043 Nucleic acid sequence Proteins 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 238000012408 PCR amplification Methods 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 6
- 102100023152 Scinderin Human genes 0.000 description 6
- 238000013461 design Methods 0.000 description 6
- 108090000765 processed proteins & peptides Proteins 0.000 description 6
- 108010073419 scinderin Proteins 0.000 description 6
- 238000001890 transfection Methods 0.000 description 6
- 102000003960 Ligases Human genes 0.000 description 4
- 108090000364 Ligases Proteins 0.000 description 4
- 108010017070 Zinc Finger Nucleases Proteins 0.000 description 4
- 238000000246 agarose gel electrophoresis Methods 0.000 description 4
- 230000004087 circulation Effects 0.000 description 4
- 238000013016 damping Methods 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 230000014509 gene expression Effects 0.000 description 4
- 101100148606 Caenorhabditis elegans pst-1 gene Proteins 0.000 description 3
- 108010019160 Pancreatin Proteins 0.000 description 3
- 238000012217 deletion Methods 0.000 description 3
- 230000037430 deletion Effects 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 239000003292 glue Substances 0.000 description 3
- 210000003128 head Anatomy 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000032965 negative regulation of cell volume Effects 0.000 description 3
- 229940055695 pancreatin Drugs 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 102100031780 Endonuclease Human genes 0.000 description 2
- 108010042407 Endonucleases Proteins 0.000 description 2
- 101000729271 Homo sapiens Retinoid isomerohydrolase Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102100031176 Retinoid isomerohydrolase Human genes 0.000 description 2
- 108091023040 Transcription factor Proteins 0.000 description 2
- 102000040945 Transcription factor Human genes 0.000 description 2
- 101710185494 Zinc finger protein Proteins 0.000 description 2
- 102100023597 Zinc finger protein 816 Human genes 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 210000004899 c-terminal region Anatomy 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 230000006801 homologous recombination Effects 0.000 description 2
- 238000002744 homologous recombination Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 210000003583 retinal pigment epithelium Anatomy 0.000 description 2
- 239000011435 rock Substances 0.000 description 2
- NCYCYZXNIZJOKI-IOUUIBBYSA-N 11-cis-retinal Chemical compound O=C/C=C(\C)/C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-IOUUIBBYSA-N 0.000 description 1
- 201000004569 Blindness Diseases 0.000 description 1
- 101150017501 CCR5 gene Proteins 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 230000007023 DNA restriction-modification system Effects 0.000 description 1
- 230000004568 DNA-binding Effects 0.000 description 1
- 101000634196 Homo sapiens Neurotrophin-3 Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 241000984622 Leucodon Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 102100029268 Neurotrophin-3 Human genes 0.000 description 1
- 238000009004 PCR Kit Methods 0.000 description 1
- 108091000054 Prion Proteins 0.000 description 1
- NCYCYZXNIZJOKI-OVSJKPMPSA-N Retinaldehyde Chemical compound O=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-OVSJKPMPSA-N 0.000 description 1
- 208000007014 Retinitis pigmentosa Diseases 0.000 description 1
- 102000004330 Rhodopsin Human genes 0.000 description 1
- 108090000820 Rhodopsin Proteins 0.000 description 1
- 108010073062 Transcription Activator-Like Effectors Proteins 0.000 description 1
- 241000589634 Xanthomonas Species 0.000 description 1
- 238000005267 amalgamation Methods 0.000 description 1
- 206010001902 amaurosis Diseases 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 230000005782 double-strand break Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 238000013412 genome amplification Methods 0.000 description 1
- 210000004263 induced pluripotent stem cell Anatomy 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 108010082117 matrigel Proteins 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- LCCNCVORNKJIRZ-UHFFFAOYSA-N parathion Chemical compound CCOP(=S)(OCC)OC1=CC=C([N+]([O-])=O)C=C1 LCCNCVORNKJIRZ-UHFFFAOYSA-N 0.000 description 1
- 230000003950 pathogenic mechanism Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 108010054624 red fluorescent protein Proteins 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 235000020945 retinal Nutrition 0.000 description 1
- 239000011604 retinal Substances 0.000 description 1
- 210000000880 retinal rod photoreceptor cell Anatomy 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000012096 transfection reagent Substances 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
TALE名称 | 靶序列 |
RPE65-TALE-L1(SEQ ID NO:16) | tccttcttcattct |
RPE65-TALE-L2(SEQ ID NO:17) | gagaacttccttcttcatt |
RPE65-TALE-L3(SEQ ID NO:18) | gagaacttccttcttcat |
RPE65-TALE-L4(SEQ ID NO:19) | gagaacttccttctt |
RPE65-TALE-R1(SEQ ID NO:20) | tcaggatccagagttct |
RPE65-TALE-R2(SEQ ID NO:21) | tcaggatccagagtt |
RPE65-TALE-R3(SEQ ID NO:22) | tcaggatccagagt |
引物名称 | 引物序列 |
F-assem(SEQ ID NO:45) | CGGGAGCCGACGTCGACAG |
R-assem(SEQ ID NO:46) | CGCTCGAGCGACACGCAGG |
引物名称 | 引物序列 |
TALE-正向测序(SEQ ID NO:47) | CTCCCCTTCAGCTGGACAC |
TALE-反向测序(SEQ ID NO:48) | AGCTGGGCCACGATTGAC |
RPE65-TALEN-R1 | RPE65-TALEN-R2 | RPE65-TALEN-R3 | |
RPE65-TALEN-L1 | L1+R1 | L1+R2 | L1+R3 |
RPE65-TALEN-L2 | L2+R1 | L2+R2 | L2+R3 |
RPE65-TALEN-L3 | L3+R1 | L3+R2 | L3+R3 |
RPE65-TALEN-L4 | L4+R1 | L4+R2 | L4+R3 |
Claims (12)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210163377.XA CN102702332B (zh) | 2012-05-23 | 2012-05-23 | 一对转录激活子样效应因子核酸酶l1和r2及其编码基因与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210163377.XA CN102702332B (zh) | 2012-05-23 | 2012-05-23 | 一对转录激活子样效应因子核酸酶l1和r2及其编码基因与应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102702332A CN102702332A (zh) | 2012-10-03 |
CN102702332B true CN102702332B (zh) | 2014-01-08 |
Family
ID=46895423
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201210163377.XA Expired - Fee Related CN102702332B (zh) | 2012-05-23 | 2012-05-23 | 一对转录激活子样效应因子核酸酶l1和r2及其编码基因与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102702332B (zh) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103898099B (zh) * | 2012-12-25 | 2017-08-25 | 袁晶 | 一种实现基因组定点修饰的蛋白表达载体的方法 |
CN103289974A (zh) * | 2013-04-28 | 2013-09-11 | 新疆农垦科学院 | 绵羊fgf5基因定点敲除系统及其应用 |
CN104450776A (zh) * | 2014-12-11 | 2015-03-25 | 中国农业科学院棉花研究所 | 共表达talen蛋白的植物表达载体及其构建方法 |
-
2012
- 2012-05-23 CN CN201210163377.XA patent/CN102702332B/zh not_active Expired - Fee Related
Non-Patent Citations (4)
Title |
---|
De novo-engineered transcription activator-like effector (TALE) hybrid nuclease with novel DNA binding specificity creates double-strand breaks;Magdy M. Mahfouz, Lixin Li, Md. Shamimuzzaman, Anjar Wibowo;《Proceeding of the National Academy of Sciences of the United State of America》;20110208;第108卷(第6期);全文 * |
Dirk Hockemeyer, Haoyi Wang, Samira Kiani,Christine S Lai.Genetic engineering of human pluripotent cells using TALE nucleases.《Nature Biotechnology》.2011,第29卷全文. * |
Magdy M. Mahfouz, Lixin Li, Md. Shamimuzzaman, Anjar Wibowo.De novo-engineered transcription activator-like effector (TALE) hybrid nuclease with novel DNA binding specificity creates double-strand breaks.《Proceeding of the National Academy of Sciences of the United State of America》.2011,第108卷(第6期),全文. |
王昕.TALE 核酸酶介导的基因组定点修饰技术.《中国生物化学与分子生物学报》.2012,第28卷(第3期),211-216. * |
Also Published As
Publication number | Publication date |
---|---|
CN102702332A (zh) | 2012-10-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102558309B (zh) | 一对转录激活子样效应因子核酸酶及其编码基因与应用 | |
US11939606B2 (en) | CRISPR-associated (Cas) protein | |
CN113015798B (zh) | CRISPR-Cas12a酶和系统 | |
WO2018208755A1 (en) | Compositions and methods for tagging target proteins in proximity to a nucleotide sequence of interest | |
CN106086031B (zh) | 猪肌抑素基因编辑位点及其应用 | |
CN109415729A (zh) | 具有降低毒性的基因编辑试剂 | |
CN109136248A (zh) | 多靶点编辑载体及其构建方法和应用 | |
CN103388006A (zh) | 一种基因定点突变的构建方法 | |
CN103820452B (zh) | 一种sgRNA片段及其应用 | |
CN112105728A (zh) | CRISPR/Cas效应蛋白及系统 | |
CN102850444A (zh) | 一对转录激活子样效应因子核酸酶l3和r1及其编码基因与应用 | |
CN111575319B (zh) | 一种高效的crispr rnp和供体dna共位介导的基因插入或替换方法及其应用 | |
CN102702332B (zh) | 一对转录激活子样效应因子核酸酶l1和r2及其编码基因与应用 | |
CN102702335B (zh) | 重组转录激活子样效应因子、转录激活子样效应因子核酸酶及其编码基因及应用 | |
CN106754949B (zh) | 猪肌抑素基因编辑位点864-883及其应用 | |
CN117187220A (zh) | 腺嘌呤脱氨酶及其在碱基编辑中的用途 | |
CA3236802A1 (en) | Serine recombinases | |
CN105838691B (zh) | 一对转录激活子样效应因子核酸酶及其编码基因与应用 | |
CN102627692B (zh) | 一对转录激活子样效应因子核酸酶及其编码基因与应用 | |
CN102702331A (zh) | 一对转录激活子样效应因子核酸酶及其编码基因与应用 | |
JP2023542976A (ja) | カーゴヌクレオチド配列を転位させるための系および方法 | |
CN116004573B (zh) | 编辑活性提高的Cas蛋白及其应用 | |
CN102627690B (zh) | 一对转录激活子样效应因子核酸酶及其编码基因与应用 | |
CN107034234A (zh) | 一种用于敲除cho细胞中fut8和dhfr两种基因的试剂盒 | |
CN102627691B (zh) | 一对转录激活子样效应因子核酸酶及其编码基因与应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: One pair of transcription activator effect factor nucleases R1 and R2, coding gene and application thereof Effective date of registration: 20140610 Granted publication date: 20140108 Pledgee: Pudong Shanghai technology financing Company limited by guarantee Pledgor: SHANGHAI SIDANSAI BIOTECHNOLOGY CO.,LTD. Registration number: 2014310000035 |
|
PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20150615 Granted publication date: 20140108 Pledgee: Pudong Shanghai technology financing Company limited by guarantee Pledgor: SHANGHAI SIDANSAI BIOTECHNOLOGY CO.,LTD. Registration number: 2014310000035 |
|
PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: One pair of transcription activator effect factor nucleases R1 and R2, coding gene and application thereof Effective date of registration: 20150701 Granted publication date: 20140108 Pledgee: Pudong Shanghai technology financing Company limited by guarantee Pledgor: SHANGHAI SIDANSAI BIOTECHNOLOGY CO.,LTD. Registration number: 2015310000025 |
|
PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20160524 Granted publication date: 20140108 Pledgee: Pudong Shanghai technology financing Company limited by guarantee Pledgor: SHANGHAI SIDANSAI BIOTECHNOLOGY CO.,LTD. Registration number: 2015310000025 |
|
PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20211207 Address after: 201203 floor 3, building 1, No. 6055, Jinhai Road, Fengxian District, Shanghai Patentee after: Shanghai Xuxu Technology Co.,Ltd. Address before: 201203 room 640, building 1, Lane 720, Cailun Road, Zhangjiang High Tech Park, Pudong New Area, Shanghai Patentee before: SHANGHAI SIDANSAI BIOTECHNOLOGY CO.,LTD. |
|
CF01 | Termination of patent right due to non-payment of annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140108 |