Abstract
Reprogrammable CRISPR/dCas9-based recruitment of DNMT1 for site-specific DNA demethylation and gene regulation
Highlights
1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; Dear Editor, DNA methylation and demethylation play a critical role in regulating gene expression as well as developmental and pathological processes[1]
To test whether the transcription of RANKL could be regulated by the CRISPR/dCas9-R2 system, five sgRNAs were designed to target the RANKL promoter region (Fig. 1c), and the impact of individual sgRNA on RANKL transcription was assayed by quantitative RT-PCR, using total RNA extraction prepared from the HEK-293T cells 5 days post transfection
The DNMT1 is recruited by the sgRNA-R2 stemloop, and the methyltransferase activity is inhibited, resulting in the decrease of the DNA methylation level. b The RNA structure of the R2 stemloop. c Illustration of the five sgRNAs designed for targeting the promoter region of the human RANKL gene
Summary
1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; Dear Editor, DNA methylation and demethylation play a critical role in regulating gene expression as well as developmental and pathological processes[1]. To test whether the transcription of RANKL could be regulated by the CRISPR/dCas9-R2 system, five sgRNAs were designed to target the RANKL promoter region (Fig. 1c), and the impact of individual sgRNA on RANKL transcription was assayed by quantitative RT-PCR (qRT-PCR), using total RNA extraction prepared from the HEK-293T cells 5 days post transfection.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
