Why the study?
Target sequence recognition by TALE is affected by mismatches, prompting the goal to construct a TALE capable of accurately distinguishing a single nucleotide difference.
The study demonstrates that introducing continuous mismatches in specific TALE repeats enables accurate discrimination and selective cleavage of DNA with a single nucleotide difference.
TALEs with single mismatches distinguish nucleotide differences; extends precision genome editing tools while leaving clinical use open.
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.
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Sakono et al. (2022) studied this question.
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