MiRNA-Responsive CRISPR-Cas System via a DNA Regulator

被引:0
|
作者
Yun, Dayoung [1 ]
Jung, Cheulhee [1 ]
机构
[1] Korea Univ, Coll Life Sci & Biotechnol, Dept Biotechnol, Seoul 02841, South Korea
来源
BIOSENSORS-BASEL | 2023年 / 13卷 / 11期
关键词
CRISPR-Cas9; DNA regulator; miRNA; specific regulation; BREAST-CANCER; MICRORNA; RNA; BIOMARKERS; PROTEIN; DISEASE; LET-7;
D O I
10.3390/bios13110975
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Clustered regularly interspaced short palindromic repeats (CRISPR)- CRISPR-associated protein 9 (Cas9) genome editing technology is widely used for gene editing because it provides versatility in genetic manipulation. Several methods for regulating CRISPR activity already exist for accurate editing, but these require complex engineering. Thus, a simple and convenient regulatory system is required. In this study, we devised a CRISPR activation system using a DNA regulator that can be activated by miRNAs. The designed regulator was divided into two parts. The inhibition component consisted of the protospacer-adjacent motif (PAM) and seed sequence, which are important for Cas9 target recognition and bind to the ribonucleoprotein (RNP) complex for inhibition. The miRNA recognition component has a single-stranded toehold DNA for target miRNA binding and a partial double-stranded DNA complementary to the remaining miRNA sequence. In the presence of target miRNAs, the structure of the regulator is disrupted by the miRNAs, leading to its dissociation from the RNP complex and subsequent restoration of CRISPR activity. This method is easy to design and can be applied to various miRNAs via simple sequence manipulation. Therefore, this strategy provides a general platform for controlled genome editing.
引用
收藏
页数:12
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