TY - JOUR
T1 - Development of a Transcriptional Activator-Like Effector Protein to Accurately Discriminate Single Nucleotide Difference
AU - Sakono, Masafumi
AU - Oya, Takuma
AU - Aoki, Mio
N1 - Publisher Copyright:
© 2022 Wiley-VCH GmbH.
PY - 2023/2/1
Y1 - 2023/2/1
N2 - 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.
AB - 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.
KW - DNA binding protein
KW - KRAS
KW - bioluminescence resonance energy transfer
KW - single nucleotide difference
KW - transcriptional activator-like effector protein
UR - http://www.scopus.com/inward/record.url?scp=85143542819&partnerID=8YFLogxK
U2 - 10.1002/cbic.202200486
DO - 10.1002/cbic.202200486
M3 - 学術論文
C2 - 36409599
AN - SCOPUS:85143542819
SN - 1439-4227
VL - 24
JO - ChemBioChem
JF - ChemBioChem
IS - 3
M1 - e202200486
ER -