RU2015111840A - Конструируемые белки с цинковыми пальцами, направленные на гены растений, вовлеченные в биосинтез жирных кислот - Google Patents
Конструируемые белки с цинковыми пальцами, направленные на гены растений, вовлеченные в биосинтез жирных кислот Download PDFInfo
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Abstract
1. Неприродный белок с цинковыми пальцами, модулирующий экспрессию по меньшей мере одного эндогенного гена тиоэстеразы жирных кислот A (FatA) или тиоэстеразы жирных кислот B (FatB) растений, где белок с цинковыми пальцами связывается с участком-мишенью, содержащим по меньшей мере два подучастка-мишени как представлено в таблице 10A (SEQ ID NO:45-72).2. Слитый белок, содержащий белок с цинковыми пальцами по п. 1 и функциональный домен.3. Слитый белок по п. 2, где функциональный домен представляет собой домен регуляции транскрипции или домен расщепления.4. Белок с цинковыми пальцами или слитый белок по любому из пп. 1-3, где ген растения содержится в растении Brassica.5. Белок с цинковыми пальцами по п. 4, где белок с цинковыми пальцами связывается с участком-мишенью, как представлено в таблице 10A.6. Белок с цинковыми пальцами по п. 4, где белок с цинковыми пальцами содержит области распознающих спиралей, представленные в строках таблицы 10B.7. Полинуклеотид, кодирующий один или более белков с цинковыми пальцами или слитых белков по любому из пп. 1-6.8. Способ модификации одного или более генов, вовлеченных в биосинтез жирных кислот в растительной клетке, где способ включает:введение в растительную клетку одного или более экспрессирующих векторов, содержащих по меньшей мере одинполинуклеотид по п. 7, в таких условиях, что экспрессируются один или более белков с цинковыми пальцами, и подвергаются модификации один из генов Fat.9. Способ по п. 8, где модификация включает модуляцию экспрессии по меньшей мере одного гена.10. Способ по п. 9, где активирована экспрессия по меньшей мере одного гена.11. Способ по п. 9 или 10, где репрессирована экспрессия по меньшей мер
Claims (18)
1. Неприродный белок с цинковыми пальцами, модулирующий экспрессию по меньшей мере одного эндогенного гена тиоэстеразы жирных кислот A (FatA) или тиоэстеразы жирных кислот B (FatB) растений, где белок с цинковыми пальцами связывается с участком-мишенью, содержащим по меньшей мере два подучастка-мишени как представлено в таблице 10A (SEQ ID NO:45-72).
2. Слитый белок, содержащий белок с цинковыми пальцами по п. 1 и функциональный домен.
3. Слитый белок по п. 2, где функциональный домен представляет собой домен регуляции транскрипции или домен расщепления.
4. Белок с цинковыми пальцами или слитый белок по любому из пп. 1-3, где ген растения содержится в растении Brassica.
5. Белок с цинковыми пальцами по п. 4, где белок с цинковыми пальцами связывается с участком-мишенью, как представлено в таблице 10A.
6. Белок с цинковыми пальцами по п. 4, где белок с цинковыми пальцами содержит области распознающих спиралей, представленные в строках таблицы 10B.
7. Полинуклеотид, кодирующий один или более белков с цинковыми пальцами или слитых белков по любому из пп. 1-6.
8. Способ модификации одного или более генов, вовлеченных в биосинтез жирных кислот в растительной клетке, где способ включает:
введение в растительную клетку одного или более экспрессирующих векторов, содержащих по меньшей мере один
полинуклеотид по п. 7, в таких условиях, что экспрессируются один или более белков с цинковыми пальцами, и подвергаются модификации один из генов Fat.
9. Способ по п. 8, где модификация включает модуляцию экспрессии по меньшей мере одного гена.
10. Способ по п. 9, где активирована экспрессия по меньшей мере одного гена.
11. Способ по п. 9 или 10, где репрессирована экспрессия по меньшей мере одного гена.
12. Способ по п. 8, где полинуклеотид кодирует нуклеазы с цинковыми пальцами и расщепляется по меньшей мере один ген.
13. Способ по п. 12, дополнительно включающий этап введения донорной нуклеиновой кислоты так, что донорный вектор посредством гомологичной рекомбинации встраивается в участок расщепления.
14. Растительная клетка, содержащая по меньшей мере один ген Fat, модифицированный любым из способов по любому из пп. 8-13.
15. Растительная клетка по п. 14, где клетка находится в семени и содержание жирных кислот в семени модифицировано.
16. Растение, содержащее по меньшей мере одну клетку по любому из пп. 14-15.
17. Семя растения по п. 16.
18. Потомство растения по п. 16.
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