ME02836B - Postupci i sastavi za rnk-usmerenu modifikaciju ciljane dnk i za rnk-usmerenu modulaciju transkripcije - Google Patents

Postupci i sastavi za rnk-usmerenu modifikaciju ciljane dnk i za rnk-usmerenu modulaciju transkripcije

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Publication number
ME02836B
ME02836B MEP-2017-180A MEP2017180A ME02836B ME 02836 B ME02836 B ME 02836B ME P2017180 A MEP2017180 A ME P2017180A ME 02836 B ME02836 B ME 02836B
Authority
ME
Montenegro
Prior art keywords
dna
rna
targeting
protein
cas9 polypeptide
Prior art date
Application number
MEP-2017-180A
Other languages
German (de)
English (en)
French (fr)
Inventor
Martin Jinek
Cate James Harrison Doudna
Wendell Lim
Lei Qi
Emmanuelle Charpentier
Krysztof Chylinski
Jennifer A Doudna
Original Assignee
Univ California
Univ Vienna
Emmanuelle Charpentier
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=49624232&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=ME02836(B) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Univ California, Univ Vienna, Emmanuelle Charpentier filed Critical Univ California
Publication of ME02836B publication Critical patent/ME02836B/me

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Claims (23)

1. Postupak modifikovanja ciljane DNK, koji postupak obuhvata dovođenje u dodir ciljane DNK sa kompleksom koji obuhvata: (a) Cas9 polipeptid i (b) pojedinačni molekul (i) RNK koja cilja DNK koja obuhvata: DNK-ciljajući segment koji obuhvata nukleotidnu sekvencu koja je komplementarna sekvenci u ciljanoj DNK, i (i) segment za vezivanje proteina koji interakuje sa navedenim Cas9 polipeptidom, gde segment za vezivanje proteina obuhvata dva komplementarna predela nukleotida koji hibridizuju kako bi formirali dvostruki RNK (dsRNK) dupleks, gde su dva navedena komplementarna predela nukleotida kovalentno povezana intervenišućim nukleotidima, gde je navedeno dovođenje u dodir in vitro ili u ćeliji ex vivo; i gde je navedeno modifikovanje cepanje ciljane DNK.
2. Postupak iz patentnog zahteva 1, gde navedeni dsRNK dupleks ima dužinu od 8 baznih parova (bp) do 30 bp.
3. Postupak iz patentnog zahteva 1 ili 2, gde je procenat komplementarnosti između nukleotida koji hibridizuju kako bi formirali dsRNK dupleks segmenta za vezivanje proteina veći od 70%.
4. Postupak iz patentnog zahteva 1, 2 ili 3, gde je ciljana DNK prisutna u bakterijskoj ćeliji, arhealnoj ćeliji, jednoćelijskom eukariotskom organizmu, ćeliji biljke, ćeliji beskičmenjaka, ili ćeliji kičmenjaka.
5. Postupak patentnog zahteva 1, 2 ili 3, gde je ciljana DNK hromozomna DNK.
6. Postupak iz bilo kog od prethodnih patentnih zahteva, gde dovođenje u dodir obuhvata uvođenje u ćeliju (a) navedenog Cas9 polipeptida, ili polinukleotida koji kodira navedeni Cas9 polipeptid, i (b) navedene RNK koja cilja DNK, ili DNK polinukleotida koji kodira navedenu RNK koja cilja DNK.
7. Postupak iz patentnog zahteva 6 gde postupak dalje obuhvata uvođenje u ćeliju donorskog polinukleotida.
8. Postupak prema bilo kom od prethodnih patentnih zahteva, gde je transdukcioni domen proteina kovalentno vezan za amino terminus Cas9 polipeptida, gde navedeni transdukcioni domen proteina omogućava prelazak Cas9 polipeptida od cistola do organele unutar ćelije.
9. Postupak prema bilo kom patentnom zahtevu 1-7 gde je transdukcioni domen proteina kovalentno vezan za karboksilni terminus Cas9 polipeptida, gde navedeni transdukcioni domen proteina omogućava prelazak Cas9 polipeptida od cistola do organele unutar ćelije.
10. Sastav koja obuhvata:(a) Cas9 polipeptid, ili polinukleotid koji kodira navedeni Cas9 polipeptid, i(b) jednomolekulska RNK koja cilja DNK, ili DNK polinukleotida koji kodira navedenu RNK koja cilja DNK, gde navedenu jednomolekulsku RNK koja cilja DNK obuhvata:(i) DNK-ciljajući segment koji obuhvata nukleotidnu sekvencu koja je komplementarna sekvenci u ciljanoj DNK, i(ii) segment za vezivanje proteina koji interakuje sa navedenim Cas9 polipeptid, gde segment za vezivanje proteina obuhvata dva komplementarna predela nukleotida koji hibridizuju kako bi formirali dvostruki RNK (dsRNK) dupleks, igde su dva navedena komplementarna predela nukleotida kovalentno povezana intervenišućim nukleotidima.
11. Sastav iz patentnog zahteva 10, gde je transdukcioni domen proteina kovalentno vezan za amino terminus Cas9 polipeptida, gde navedeni transdukcioni domen proteina omogućava prelazak Cas9 polipeptida od cistola do organele unutar ćelije.
12. Sastav iz patentnog zahteva 10, gde je transdukcioni domen proteina kovalentno vezan za karboksilni terminus Cas9 polipeptida, gde navedeni transdukcioni domen proteina omogućava prelazak Cas9 polipeptida od cistola do organele unutar ćelije.
13. Sastav bilo kog patentnog zahteva 10-12, gde (a) Cas9 polipeptid, ili polinukleotid koji kodira navedeni Cas9 polipeptid, i (b) jednomolekulska RNK koja cilja DNK, ili DNK polinukleotida koji kodira navedenu RNK koja cilja DNK su u ćeliji in vitro ili ex vivo, gde ćelija nije ljudska germ ćelija.
14. Jednomolekulska RNK koja cilja DNK, ili DNK polinukleotida koji kodira navedenu RNK koja cilja DNK, gde jednomolekulska RNK koja cilja DNK obuhvata:(a) DNK-ciljajući segment koji obuhvata nukleotidnu sekvencu koja je komplementarna ciljanoj sekvenci ciljane DNK, i(b) segment za vezivanje proteina koji interakuje sa Cas9 proteinom, gde segment za vezivanje proteina obuhvata dva komplementarna predela nukleotida koji hibridizuju kako bi formirali dvostruki RNK (dsRNK) dupleks, i gde su dva navedena komplementarna predela nukleotida kovalentno povezana intervenišućim nukleotidima.
15. Jedna ili više nukleinska kiselina koja obuhvata:(a) prvu nukleotidnu sekvencu koja kodira jednomolekulska RNK koja cilja DNK koja obuhvata:(i) DNK-ciljajući segment koji obuhvata nukleotidnu sekvencu koja je komplementarna ciljanoj sekvenci ciljanoj DNK, i(ii) segment za vezivanje proteina koji interakuje sa Cas9 polipeptidom, gde segment za vezivanje proteina obuhvata dva komplementarna predela nukleotida koji hibridizuju kako bi formirali dvostruki RNK (dsRNK) dupleks, i gde dva navedena komplementarna predela nukleotida kovalentno povezana intervenišućim nukleotidima;gde je prva nukleotidna sekvenca koja kodira navedenu RNK koja cilja DNK operativno povezana za promoter; i, opciono(b) drugu nukleotidnu sekvencu koja kodira Cas9 polipeptid, gde je nukleotidna sekvenca koja kodira navedeni Cas9 polipeptid operativno povezana za promoter.
16. Jedna ili više nukleinska kiselina iz patentnog zahteva 15, gde je navedena nukleinska kiselina jedan ili više rekombinantni vektor eksprimiranja.
17. Jedna ili više nukleinska kiselina iz patentnog zahteva 15, gde su jedan ili više rekombinantni vektor eksprimiranja virusni vektori.
18. Komplet koji obuhvata(a) Cas9 polipeptid, ili nukleinska kiselina koja obuhvata nukleotidnu sekvencu koja kodira navedeni Cas9 polipeptid; i(b) jednomolekulsku RNK koja cilja DNK, ili nukleinsku kiselinu koja obuhvata nukleotidnu sekvencu koja kodira navedenu jednomolekulsku RNK koja cilja DNK, gde jednomolekulska RNK koja cilja DNK obuhvata:(i) DNK-ciljajući segment koji obuhvata nukleotidnu sekvencu koja je komplementarna sekvenci u ciljanoj DNK, i(ii) segment za vezivanje proteina koji interakuje sa navedenim Cas9 polipeptidom, gde segment za vezivanje proteina obuhvata dva komplementarna predela nukleotida koji hibridizuju kako bi formirali dvostruki RNK (dsRNK) dupleks, i gde su dva navedena komplementarna predela nukleotida kovalentno povezana intervenišućim nukleotidima, igde su (a) i (b) u istom ili u različitim kontejnerima.
19. Sastav bilo kog patentnog zahteva 10-13, jednomolekulska RNK koja cilja DNK ili polinukleotid koji kodira navedenu jednomolekulsku RNK koja cilja DNK patentnog zahteva 14, jedna ili više nukleinska kiselina bilo kog patentnog zahteva 15-17 ili komplet patentnog zahteva 18, gde navedeni dsRNK dupleks ima dužinu od 8 baznih parova (bp) do 30 bp.
20. Sastav bilo kog patentnog zahteva 10-13 ili 19, jednomolekulska RNK koja cilja DNK ili polinukleotid koji kodira navedenu jednomolekulsku RNK koja cilja DNK patentnog zahteva 14 ili 19, jedna ili više nukleinska kiselina bilo kog patentnog zahteva 15-17 ili 19, ili komplet patentnog zahteva 18 ili 19, gde je procenat komplementarnosti između nukleotida koji hibridizuju kako bi formirali dsRNK dupleks segmenta za vezivanje proteina veći od 70%.
21. Sastav bilo kog patentnog zahteva 10-13, 19 ili 20, jednomolekulska RNK koja cilja DNK ili polinukleotid koji kodira navedenu jednomolekulsku RNK koja cilja DNK bilo kog patentnog zahteva 14, 19 ili 20, jedna ili više nukleinska kiselina bilo kog patentnog zahteva 15-17, 19 ili 20, ili komplet bilo kog patentnog zahteva 18, 19 ili 20, gde je ciljana DNK prisutna u bakterijskoj ćeliji, arhealnoj ćeliji, jednoćelijskom eukariotskom organizmu, ćeliji biljke, ćeliji beskičmenjaka, ili ćeliji kičmenjaka.
22. Sastav bilo kog patentnog zahteva 10-13 ili 19-21, jednomolekulska RNK koja cilja DNK ili polinukleotid koji kodira navedenu jednomolekulsku RNK koja cilja DNK bilo kog patentnog zahteva 14 ili 19-21, jedna ili više nukleinska kiselina bilo kog patentnog zahteva 15-17 ili 19-21, ili komplet bilo kog patentnog zahteva 18-21, gde je ciljana DNK hromozomna DNK.
23. Sastav bilo kog patentnog zahteva 10-13 ili 19-22, ili jednomolekulska RNK koja cilja DNK ili polinukleotid koji kodira navedenu jednomolekulsku RNK koja cilja DNK bilo kog patentnog zahteva 14 ili 19-22, ili jedna ili više nukleinska kiselina bilo kog patentnog zahteva 15-17 ili 19-22, za primenu u terapeutskom tretmanu pacijenta.
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US20190002922A1 (en) 2019-01-03
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