CRISPR/Cas9-mediated knockout of factors in non-homologous end joining pathway enhances gene targeting in silkworm cells

Li Zhu, Hiroaki Mon, Jian Xu, Man Lee, Takahiro Kusakabe

研究成果: ジャーナルへの寄稿記事

15 引用 (Scopus)

抄録

Gene targeting can be achieved by precise genetic modifications through homology-directed repair (HDR) after DNA breaks introduced by genome editing tools such as CRISPR/Cas9 system. The most common form of HDR is homologous recombination (HR). Binding to the DNA breaks by HR factors is thought to compete with non-homologous end joining (NHEJ), an alternative DNA repair pathway. Here, we knocked out the factors in NHEJ by CRISPR/Cas9 system in silkworm cells, so that increased the activities of HR up to 7-fold. Also efficient HR-mediated genome editing events occurred between the chromosomal BmTUDOR-SN gene and donor DNA sequences with an EGFP gene in the middle of two homologous arms for the target gene. Utilizing the NHEJ-deficient silkworm cells, we found that homologous arms as short as 100 bp in donor DNA could be designed to perform precise genome editing. These studies should greatly accelerate investigations into genome editing of silkworm.

元の言語英語
記事番号18103
ジャーナルScientific reports
5
DOI
出版物ステータス出版済み - 12 10 2015

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Clustered Regularly Interspaced Short Palindromic Repeats
Bombyx
Gene Targeting
Homologous Recombination
DNA Breaks
Genes
Recombinational DNA Repair
DNA Repair
Gene Editing
DNA

All Science Journal Classification (ASJC) codes

  • General

これを引用

CRISPR/Cas9-mediated knockout of factors in non-homologous end joining pathway enhances gene targeting in silkworm cells. / Zhu, Li; Mon, Hiroaki; Xu, Jian; Lee, Man; Kusakabe, Takahiro.

:: Scientific reports, 巻 5, 18103, 10.12.2015.

研究成果: ジャーナルへの寄稿記事

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