Diphtheria toxin-mediated transposon-driven poly (A)-trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells

Bai, Jie; Kondo, Ryohei; Mayasari, N. Ika et al.

Genesis, 2020, 58(9), e23386

Number of Access:583（2025-08-22 01:06 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/78542

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Title
Diphtheria toxin-mediated transposon-driven poly (A)-trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells
Creator
NRID
1000060775336 1000060775336
CreatorName
Bai, Jie
CreatorName
Kondo, Ryohei
CreatorName
Mayasari, N. Ika
NRID
1000040457148 1000040457148
CreatorName
Shigeoka, Toshiaki
NRID
1000020444523 1000020444523
CreatorName
Isotani, Ayako
NRID
1000020304066 1000020304066
ORCID
0000-0001-9859-6217 0000-0001-9859-6217
researchmap permalink
read0054910 read0054910
Scopus Author ID
16689054500 16689054500
CreatorName
Ikawa, Masahito
NRID
1000070349447 1000070349447
CreatorName
Sashida, Goro
NRID
1000000195041 1000000195041
CreatorName
Kawaichi, Masashi
NRID
1000010221756 1000010221756
CreatorName
Ishida, Yasumasa
Subject
cell lineage ablation
diphtheria toxin
embryonic stem cells
gene trap
Tol2 transposon
Description
Bai, J, Kondo, R, Mayasari, NI, et al. Diphtheria toxin‐mediated transposon‐driven poly (A)‐trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells. genesis. 2020; 58:e23386. https://doi.org/10.1002/dvg.23386
Abstract
Random gene trapping is the application of insertional mutagenesis techniques that are conventionally used to inactivate protein-coding genes in mouse embryonic stem (ES) cells. Transcriptionally silent genes are not effectively targeted by conventional random gene trapping techniques, thus we herein developed an unbiased poly (A) trap (UPATrap) method using a Tol2 transposon, which preferentially integrated into active genes rather than silent genes in ES cells. To achieve efficient trapping at transcriptionally silent genes using random insertional mutagenesis in ES cells, we generated a new diphtheria toxin (DT)-mediated trapping vector, DTrap that removed cells, through the expression of DT that was induced by the promoter activity of the trapped genes, and selected trapped clones using the neomycin-resistance gene of the vector. We found that a double-DT, the dDT vector, dominantly induced the disruption of silent genes, but not active genes, and showed more stable integration in ES cells than the UPATrap vector. The dDT vector disrupted differentiated cell lineage genes, which were silent in ES cells, and labeled trapped clone cells by the expression of EGFP upon differentiation. Thus, the dDT vector provides a systematic approach to disrupt silent genes and examine the cellular functions of trapped genes in the differentiation of target cells and development.
Publisher
John Wiley and Sons Inc
Source Title
Genesis
Volume (Issue)
58(9)
Page
e23386
Date of Issued
2020-09-16
Language
English
Handle URL
https://hdl.handle.net/11094/78542
PISSN
1526954X
eISSN
1526968X
NCID
AA11620250
Relation.isIdenticalTo
DOI (Publisher Version)
https://doi.org/10.1002/dvg.23386
PMID
32645254
Access Rights
open access
CopyRight
© 2020 The Authors. Genesis published by Wiley Periodicals LLC. This article is licensed under a Creative Commons Attribution 4.0 International License.
oaire:version
Version of Record
Category
学術雑誌論文 Journal Article