LV10655B - Endogenous gene expression modification with regulatory element - Google Patents
Endogenous gene expression modification with regulatory element Download PDFInfo
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- LV10655B LV10655B LVP-93-27A LV930027A LV10655B LV 10655 B LV10655 B LV 10655B LV 930027 A LV930027 A LV 930027A LV 10655 B LV10655 B LV 10655B
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- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/90—Stable introduction of foreign DNA into chromosome
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/67—General methods for enhancing the expression
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- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
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- C12N2830/001—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination
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- C12N2840/00—Vectors comprising a special translation-regulating system
- C12N2840/20—Vectors comprising a special translation-regulating system translation of more than one cistron
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Claims (27)
- LV 10655 Fatentformulas punkti 1. Paņēmiens parasti transkripcijā klusējoša gēna aktivēšanai šūnu līnijas genomā lai dotu iespēju minētajai līnijai transkribēt minētā gēna produktu, kas atšķiras ar to, ka minētajā genomā ar homologo rekombināciju iestarpina DNS veidojumu, pie kam minētais veidojums satur regulējošu DNS segmentu, kurš spēj stimulēt minētā gēna ekspresiju, kad ir tam operatīvi pievienots, un mērķa DNS segmentu, kurš minētajā genomā atrodas minētajā gēnā vai tā tuvumā; pie tam minētais DNS veidojums tiek iestarpināts tādā veidā, ka minētais regulējošais segments ir operatīvi saistīts ar aktivējamo gēnu.
- 2. Paņēmiens gēna ekspresijas raksturojuma modifikācijai šūnu līnijas genomā, kas atšķiras ar to, ka minētajā genomā ar homologo rekombināciju iestarpina DNS veidojumu, kas satur: regulējo šu DNS segmentu, kurš spēj modificēt minētā gēna ekspresijas raksturojumu, kad ir ar to operatīvi saistīts minētā gēna esošā regulējošā DNS segmenta vietā; mērķa DNS segmentu, kurš atrodas minētajā genomā minētajā gēnā vai tā tuvumā; pie kam minētais veidojums tiek iestarpināts tāda veida, ka minētais regulējošais segments ir operatīvi t 1 saistīts ar aktivejamo gēnu.
- 3. Paņēmiens gēna ekspresijas raksturojuma modifikācijai šūnu līnijas genomā, kas atšķiras ar to, ka minētajā genomā ar homologo rekombināciju iestarpina DNS veidojumu, kurš satur: ekspre-sējamu, amplificējamu gēnu, kurš ir spējīgs amplifioēt minēto gēnu, kad ir iestarpināts tam pietiekoši tuvu; mērķa DKS segnientu, kurš minētajā genomā atrodas minētajā gēnā vai tā tuvumā; pie tam minētais veidojums tiek iestarpināts tā, ka minētais amplifioējamais gēns atrodas pietiekoši tuvu minētajam aktivējamajam gēnam lai izsauktu tā amplifikāciju, kad tiek amplificēts minētais amplifioējamais gēne.
- 4. Paņēmiens pēc punkta 3, kas atSUras ar to, ka minētais DKS veidojums papildus satur vismaz vienu ekspresējamu izvēles mārkergēnu, kurš ir novietots tā, lai varētu tikt iestarpināts ar minēto ekspresējamajo un amplifioējamajo gēnu.
- 5. Paņēmiens pēc punkta 1, 2 vai 3, kas atšķiras ar to, ka minētais DKS veidojums satur divus mērķa segmentus, kuri abi atrodas minētajā genomā minētajā gēnā vai tā tuvumā, pie tam viens no merka segmentiem atrodas pirms minēta regulējoša segmenta, bet otrs merka segments atrodas peo minēta regulējošā segmenta.
- 6. Paņēmiens pēo punkta 1, 2 vai 3, kas atšķiras ar to, ka minētais DKS veidojums papildus satur vismaz vienu ekspresējamu izvēles mārkergēnu, kas novietots tā, ka var tikt iestarpināts ar minēto regulējošo segmentu. 2 LV 10655
- 7. Paņēmiens pēo jebkura no punktiem 1-6, kaB atšķiras ar to, ka minētais DKS veidojums papildus satur negatīvas izvēles mārkergēnu, kas novietots attiecībā pret minēto mērķa segmentu tā, ka netiek iestarpināts kopā ar minēto veidojumu tā pareizas iestarpinā-šanas procesā ar homologo rekombināciju un netiek arī ekspresēts šūnās, kurās minētais DHS veidojums tiek pareizi iestarpināts.
- 8. Paņēmiens pēo jebkura no punktiem 1, 2, 5, 6 vai 7, t kas atšķiras ar to, ka minētais DHS veidojums papildus satur ekspresijamu un amplifioējamu gēnu, kurš novietots tā, ka tiek iestarpināts ar minēto regulējošo segmentu.
- 9. Paņēmiens pēc jebkura no punktiem 1, 2, 3, 4, 5, 6, 7 vai 8, kas atšķiras ar to, ka minētā šūnu līnija ir eukariotisku šūnu līnija.
- 10. Paņēmiens pēo punkta 9, kas atšķiras ar to, ka minētā šūnu līnija ir dzīvnieku šūnu līnija.
- 11. Paņēmiens pēo punkta 9, kas atšķiras ar to, ka minētā šūnu līnija ir zīdītāju šūnu līnija.
- 12. Paņēmiens pēc punkta 9, kas atšķiras ar to, ka minētā šūnu līnija ir augu šūnu līnija.
- 13. Paņēmiens pēo punkta 6, kas atšķiras ar to, ka papildus minētajam, lai nodrošinātu minētā gēna produkta ekspresiju, 3 pēo minētās iestarpināšanas veio: minētās šūnu līnijas klonu selekoiju, kuri ekspresē minētā izvēles mārkergēna produktu; izvēlēto klonu kultivēšanu apstākļos, kuri nodrošina minētā gēna produkta ekspresiju; minētā gēna produkta savākšanu.
- 14. Paņēmiens pēo punkta 13, kaB atšķiras ar to, ka minētais izvēles mārkergēns ir neomioīna reziBtenoes gēns un minētais selekoijas posms ietver to klonu selekoiju, kuri satur neomioīna re-zistenoes gēnu. i·
- 15. Paņēmiens pēo punkta 13 vai14 , kas atšķiras ar to, ka minētais DNS veidojums papildus satur negatīvas izvēleB mārkergē-nu, kurš novietots attiecībā pret minēto mārka eegmentu tā, ka tas netiek iestarpināts kopā ar minēto veidojumu tā pareizae iestarpinā-šanas prooesā ar homologo rekombināoiju un netiek arī ekspresētB šūnās, kurās minētais DUS veidojums tiek pareizi iestarpināts, un minētais selekoijas posms ietver to klonu selekoiju, kuri nee. kspresē minēto negatīvās izvēles gēnu.
- 16. Paņēmiens pēo punkta 15, kas atšķiras ar to, ka minētais negatīvās izvēles mārkergēns ir Kerpes Simplex vīrusa timi-dīna kināzes gēnB, un minētais selekoijas posms ietver to klonu se-lekoiju, kuri iztur ekspozīciju vidē, kura nonāvē šūnaB, kurās notiek minētā gēna ekepreBija. 4 LV 10655
- 17. Šūnu līnijas genoms ar dabiskajam gēnam segmenta iestarpinā-šanas vietā operatīvi pievienotu regulējošu DNS segmentu, kas atšķiras ar noteiktu DNS secību, pie kam minētais regulējošais DNS segments dabīgā gēnā vietā neatrodas minētajā vietā.
- 18. Šūnu līnija, kas spēj ekspresēt parasti transkripoionāli klusējoša gēna produktu minētās šuunu līnijas genomā, kas atšķiras ar to, ka minētais genoms satur tajā iestarpinātu regulējošu DNS segmentu, kas operatīvi saistīts ar minēto parasti transkripoionāli klusējošo gēnu, pie kam minētais regulējošais DNS segments spēj veipināt minētās šūnu līnijas gēna produkta ekspresiju.
- 19. Šūnu līnija, kas spēj pastiprināti ekspresēt gēna produktu salīdzinot ar šūnu līniju, no kuras tā ir izveidota, kas atšķiras ar to, ka minētais gēna produkts ir minētās šūnas genoma endogēnā gēna ekspresijas produkts un minētajā genomā atrodas operatīvā veidā minētajam endogēnajam gēnam blakus vai tā tuvumā iestarpināts eksogēns regulējošs DNS segments un/vai amplifi-oējams gēns, kas spēj pastiprināt minētā gēna produkta ekspresiju minētājā šūnu līnija.
- 20. Šūnu līnija pēo punkta 19, kas atšķiras ar to, ka tajā minētais eksogēnais regulējošais DNS segments un/vai amplifioē-jamais gēns ir eksogēns regulējošais DNS segments.
- 21. Sūnu līnija peo punkta 19, kas atšķiras ar to, ka tajā minētais eksogēnais regulējošais DNS segments un/vai arnpliiicē- 5 jamais gēns ir eksogēns amplifioē jamais gēns.
- 22. Sūnu līnija pēo punkta 18 vai 20, kas atšķiras ar to, ka minētaiB regulējošais DKS segments ir tāds, kas spēj veicināt gēna produkta ekspresiju, ko normāli ekspresē minētā šūnu līnija vai mikroorganisms.
- 23. Šūnu līnija pēo punkta 22, kae atšķiras ar to, ka iestarpinātais regulējošais DKS segments ir daļa no DKS veidojuma, kura sastāvā ir minētais regulējošais DKS segmente un vismaz viens izvēles mārkergēns.
- 24. Šūnu līnija pēo punkta 23, kas atšķiras ar to,ka minētais DKS veidojums papildus satur amplifioē jamu gēnu. !
- 25. Paņēmiens gēna produkta iegūšanai no šūnu līnijas, kas a t š k i r a b ar to, ka diferencētu šūnu līniju pēo jebkura no punktiem 15 - 24 kultivē apstākļos, kas ļauj notikt minētā gēna ekspresijai un savāo minēto gēna produktu.
- 26. DKS veidojums iestarpināšanai noteiktā saimnieka šūnu līnijā, kas atšķiras ar to, ka satur regulējošo DKS segmentu, kurš spēj modificēt gēnu ekspresijas raksturojumus saimnieka šūnu līnijā, kad tas ir operatīvi saistīts pie gēna, un DKS mērķa segmentu, kurš atrodas Baimnieka šūnu līnijae genomā izvēlētā gēna tuvumā.
- 27. DKS veidojums iestarpināšanai noteiktā saimnieka šūnu līni- 6 LV 10655 jā, kas atšķiras ar to, ka satur ekspresijamu, amplifioējamu gēnu, kurš spēj amplifioēt gēnu saimnieka šūnu līnijā, kad ir iestarpināts tam pietiekoši tuvu, un DNS mērķa segmentu, kas atrodas saimnieka šūnu līnijas genomā tuvumā izvēlētajam gēnam. »· 7
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US45478389A | 1989-12-22 | 1989-12-22 | |
PCT/US1990/007642 WO1991009955A1 (en) | 1989-12-22 | 1990-12-21 | Endogenous gene expression modification with regulatory element |
Publications (2)
Publication Number | Publication Date |
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LV10655A LV10655A (lv) | 1995-04-20 |
LV10655B true LV10655B (en) | 1995-08-20 |
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Application Number | Title | Priority Date | Filing Date |
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LVP-93-27A LV10655B (en) | 1989-12-22 | 1993-01-13 | Endogenous gene expression modification with regulatory element |
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EP (5) | EP1484412A3 (lv) |
JP (3) | JP3501286B2 (lv) |
KR (1) | KR0176693B1 (lv) |
AR (1) | AR245776A1 (lv) |
AT (2) | ATE156189T1 (lv) |
AU (1) | AU645294B2 (lv) |
BG (1) | BG60624B1 (lv) |
BR (1) | BR9007937A (lv) |
CA (1) | CA2071989C (lv) |
DE (3) | DE69031172T2 (lv) |
DK (2) | DK0505500T3 (lv) |
ES (2) | ES2104690T3 (lv) |
FI (1) | FI107391B (lv) |
GR (1) | GR3025057T3 (lv) |
HK (2) | HK1000547A1 (lv) |
HU (1) | HU217212B (lv) |
IL (4) | IL116046A (lv) |
LT (1) | LT3998B (lv) |
LV (1) | LV10655B (lv) |
NO (1) | NO311896B1 (lv) |
OA (1) | OA09594A (lv) |
RO (1) | RO109864B1 (lv) |
RU (1) | RU2128227C1 (lv) |
UA (1) | UA42675C2 (lv) |
WO (1) | WO1991009955A1 (lv) |
ZA (1) | ZA9010392B (lv) |
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