Sequence-specific retrotransposition of 28S rDNA-specific LINE R2Ol in human cells

© 2019 Su et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society..

R2 is a long interspersed element (LINE) found in a specific sequence of the 28S rDNA among a wide variety of animals. Recently, we observed that R2Ol isolated from medaka fish, Oryzias latipes, retrotransposes sequence specifically into the target sequence of zebrafish. Because the 28S target and flanking regions are widely conserved among vertebrates, we examined whether R2Ol can also integrate in a sequence-specific manner in human cells. Using adenovirus-mediated expression of R2Ol constructs, we confirmed an accurate insertion of R2Ol into the 28S target of human 293T cells. However, the R2Ol mutant devoid of endonuclease (EN) activity showed no retrotransposition ability, suggesting that the sequence-specific integration of R2Ol into 28S rDNA occurs via the cleavage activity of EN. By introducing both R2Ol helper virus and donor plasmid in human cells, we succeeded in retrotransposing an exogenous EGFP gene into the 28S target site by the trans-complementation system, which enabled simplification of specific gene knock-in in a time-efficient manner. We believe that R2Ol may provide an alternative targeted gene knock-in method for practical applications such as gene therapy in future.

Medienart:

E-Artikel

Erscheinungsjahr:

2019

Erschienen:

2019

Enthalten in:

Zur Gesamtaufnahme - volume:25

Enthalten in:

RNA (New York, N.Y.) - 25(2019), 11 vom: 03. Nov., Seite 1432-1438

Sprache:

Englisch

Beteiligte Personen:

Su, Yuting [VerfasserIn]
Nichuguti, Narisu [VerfasserIn]
Kuroki-Kami, Azusa [VerfasserIn]
Fujiwara, Haruhiko [VerfasserIn]

Links:

Volltext

Themen:

28S rDNA specific element R2Ol
DNA, Ribosomal
Gene therapy
Human cell
Journal Article
Long interspersed element (LINE)
RNA, Ribosomal, 28S
Retroelements
Sequence-specific retrotransposition
Targeted gene knock-in

Anmerkungen:

Date Completed 06.01.2020

Date Revised 06.01.2020

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1261/rna.072512.119

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM300446330