RRC ID 73455
Author Sakono M, Oya T, Aoki M.
Title Development of a Transcriptional Activator-Like Effector Protein to Accurately Discriminate Single Nucleotide Difference.
Journal Chembiochem
Abstract 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.
Pages e202200486
Published 2022-11-21
DOI 10.1002/cbic.202200486
PMID 36409599
Resource
DNA material Genome Network Project Human cDNA clone IRAK014C23 (HGX005671)