Development of a Transcriptional Activator-Like Effector Protein to Accurately Discriminate Single Nucleotide Difference

Masafumi Sakono*, Takuma Oya, Mio Aoki

*この論文の責任著者

研究成果: ジャーナルへの寄稿学術論文査読

1 被引用数 (Scopus)

抄録

Transcriptional activator-like effector (TALE), a DNA-binding protein, is widely used in genome editing. However, the recognition of the target sequence by the TALE is adversely affected by the number of mismatches. Therefore, the association constant of DNA-TALE complex formation can be controlled by appropriately introducing a mismatch into the TALE recognition sequence. This study aimed to construct a TALE that can distinguish a single nucleotide difference. Our results show that a single mismatch present in repeats 2 or 3 of TALE did not interfere with the complex formation with DNA, whereas continuous mismatches present in repeats 2 and 3 significantly reduced association with the target DNA. Based on these findings, we constructed a detection system of the one nucleotide difference in gene with high accuracy and constructed a TALE-nuclease (TALEN) that selectively cleaves DNA with a single mismatch.

本文言語英語
論文番号e202200486
ジャーナルChemBioChem
24
3
DOI
出版ステータス出版済み - 2023/02/01

ASJC Scopus 主題領域

  • 生化学
  • 分子医療
  • 分子生物学
  • 有機化学

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