Peculiarities in Creation of Genetic Engineering Constructions for Knock-In Variant of Genome Editing ofArabidopsis thalianaCell Culture

Belavin P. A.; Permyakova N. V.; Zagorskaya A. A.; Marenkova T. V.; Sidorchuk Yu. V.; Uvarova E. A.; Rozov S. M.; Deineko E. V.

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Peculiarities in Creation of Genetic Engineering Constructions for Knock-In Variant of Genome Editing ofArabidopsis thalianaCell Culture

机译：基因组植物基因组植物基因组培养基因组变种植入工程结构的特点

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摘要

The peculiarities in the creation of genetic engineering tools for theknock-invariant of genome editing are considered in detail on the example ofgfpgene delivery to two target regions (nucleolus organizer region and the region of one of histone H3 genes) of theArabidopsis thaliana(L.) Heynh. genome using different methods of delivering exogenous DNA (agrobacterium-mediated transformation, bio-ballistics using different vectors and RNP complexes). Differences in the approaches to the creation of donor constructions and Cas9 tools depending on the selected method of delivery are considered. The selected target regions are of interest for further biotechnological studies on the creation of recombinant protein bioproducer lines since they refer to "housekeeping" gene regions and are characterized by a high transcriptional activity. It was established that the selected regions are not equivalent to each other as targets for incorporation of exogenous DNA. A complex compartmentalization of nucleolus, as well as a unique mechanism of "neutralization" of double-strand breaks, acts as barriers preventing the delivery of genetic engineering tools to this region. The second region of "housekeeping" genes (histoneH3.3gene region) seems accessible and can be used for theknock-invariant of genome editing.

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来源
《Russian Journal of Plant Physiology》 |2020年第5期|共12页
作者
Belavin P. A.; Permyakova N. V.; Zagorskaya A. A.; Marenkova T. V.; Sidorchuk Yu. V.; Uvarova E. A.; Rozov S. M.; Deineko E. V.;
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作者单位

Russian Acad Sci Fed Res Ctr Inst Cytol &

Genet Siberian Branch Novosibirsk 630090 Russia;

Russian Acad Sci Fed Res Ctr Inst Cytol &

Genet Siberian Branch Novosibirsk 630090 Russia;

Russian Acad Sci Fed Res Ctr Inst Cytol &

Genet Siberian Branch Novosibirsk 630090 Russia;

Russian Acad Sci Fed Res Ctr Inst Cytol &

Genet Siberian Branch Novosibirsk 630090 Russia;

Russian Acad Sci Fed Res Ctr Inst Cytol &

Genet Siberian Branch Novosibirsk 630090 Russia;

Russian Acad Sci Fed Res Ctr Inst Cytol &

Genet Siberian Branch Novosibirsk 630090 Russia;

Russian Acad Sci Fed Res Ctr Inst Cytol &

Genet Siberian Branch Novosibirsk 630090 Russia;

Russian Acad Sci Fed Res Ctr Inst Cytol &

Genet Siberian Branch Novosibirsk 630090 Russia;

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原文格式 PDF
正文语种 eng
中图分类植物学;
关键词
Arabidopsis thaliana; genome editing; knock-in; nucleolus organizer; histone H3; 3 gene; cell culture;

机译：拟南芥天然;基因组编辑;敲入;核仁组织者;组蛋白H3;3基因;细胞培养;

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Peculiarities in Creation of Genetic Engineering Constructions for Knock-In Variant of Genome Editing ofArabidopsis thalianaCell Culture

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