Aptazyme-mediated direct modulation of post-transcriptional sgRNA level for conditional genome editing and gene expression

Hao Chen; Yanqing Li; Chunhua Du; Yan Li; Junli Zhao; Xiaojing Zheng; Qinwen Mao; Haibin Xia

doi:10.1016/j.jbiotec.2018.10.011

Aptazyme-mediated direct modulation of post-transcriptional sgRNA level for conditional genome editing and gene expression

Hao Chen, Yanqing Li, Chunhua Du, Yan Li, Junli Zhao, Xiaojing Zheng, Qinwen Mao, Haibin Xia^*

^*Corresponding author for this work

Pathology

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

RNA-guided endonuclease Cas9 derived from microbial CRISPR-Cas adaptive immune systems is a powerful tool for genome editing, which has been widely used in eukaryotic systems, prokaryotic systems, and plants. However, the off-target effects caused by Cas9/sgRNA remain a major concern. Currently, the efforts to reduce the off-target effects mainly focus on improving the targeting specificity of sgRNA/Cas9, regulating the activity of the Cas9 protein or the sgRNA, and controlling the time window of their expression. In this study, a novel system was established to regulate the post-transcriptional sgRNA level by small molecule-controlled aptazyme. This system was shown to reduce the off-target effects caused by Cas9/sgRNA, while enabling precise temporal control over gene editing and regulatory activity. This new system could provide a potentially safer and more powerful tool for genome editing and therapeutic application.

Original language	English (US)
Pages (from-to)	23-29
Number of pages	7
Journal	Journal of Biotechnology
Volume	288
DOIs	https://doi.org/10.1016/j.jbiotec.2018.10.011
State	Published - Dec 20 2018

Funding

This work was supported by the Foundation for Excellent Doctoral Degree Dissertation [ X2011YB09 ] of Shaanxi Normal University and research grants to H.X. from the National Natural Science Foundation of China [No. 81471772 No. 81773265 ] and the Key Research and Development Plan of Shaanxi Province (No. 2018SF-106 ).

Keywords

Aptazyme
Cas9
Genome editing
Off-target effect
Theophylline
sgRNA

ASJC Scopus subject areas

Applied Microbiology and Biotechnology
Bioengineering
Biotechnology

Access to Document

10.1016/j.jbiotec.2018.10.011

Cite this

@article{a618a2099fd64108a20dbcbdf22c72f3,

title = "Aptazyme-mediated direct modulation of post-transcriptional sgRNA level for conditional genome editing and gene expression",

abstract = "RNA-guided endonuclease Cas9 derived from microbial CRISPR-Cas adaptive immune systems is a powerful tool for genome editing, which has been widely used in eukaryotic systems, prokaryotic systems, and plants. However, the off-target effects caused by Cas9/sgRNA remain a major concern. Currently, the efforts to reduce the off-target effects mainly focus on improving the targeting specificity of sgRNA/Cas9, regulating the activity of the Cas9 protein or the sgRNA, and controlling the time window of their expression. In this study, a novel system was established to regulate the post-transcriptional sgRNA level by small molecule-controlled aptazyme. This system was shown to reduce the off-target effects caused by Cas9/sgRNA, while enabling precise temporal control over gene editing and regulatory activity. This new system could provide a potentially safer and more powerful tool for genome editing and therapeutic application.",

keywords = "Aptazyme, Cas9, Genome editing, Off-target effect, Theophylline, sgRNA",

author = "Hao Chen and Yanqing Li and Chunhua Du and Yan Li and Junli Zhao and Xiaojing Zheng and Qinwen Mao and Haibin Xia",

note = "Publisher Copyright: {\textcopyright} 2018 Elsevier B.V.",

year = "2018",

month = dec,

day = "20",

doi = "10.1016/j.jbiotec.2018.10.011",

language = "English (US)",

volume = "288",

pages = "23--29",

journal = "Journal of Biotechnology",

issn = "0168-1656",

publisher = "Elsevier",

}

TY - JOUR

T1 - Aptazyme-mediated direct modulation of post-transcriptional sgRNA level for conditional genome editing and gene expression

AU - Chen, Hao

AU - Li, Yanqing

AU - Du, Chunhua

AU - Li, Yan

AU - Zhao, Junli

AU - Zheng, Xiaojing

AU - Mao, Qinwen

AU - Xia, Haibin

PY - 2018/12/20

Y1 - 2018/12/20

N2 - RNA-guided endonuclease Cas9 derived from microbial CRISPR-Cas adaptive immune systems is a powerful tool for genome editing, which has been widely used in eukaryotic systems, prokaryotic systems, and plants. However, the off-target effects caused by Cas9/sgRNA remain a major concern. Currently, the efforts to reduce the off-target effects mainly focus on improving the targeting specificity of sgRNA/Cas9, regulating the activity of the Cas9 protein or the sgRNA, and controlling the time window of their expression. In this study, a novel system was established to regulate the post-transcriptional sgRNA level by small molecule-controlled aptazyme. This system was shown to reduce the off-target effects caused by Cas9/sgRNA, while enabling precise temporal control over gene editing and regulatory activity. This new system could provide a potentially safer and more powerful tool for genome editing and therapeutic application.

AB - RNA-guided endonuclease Cas9 derived from microbial CRISPR-Cas adaptive immune systems is a powerful tool for genome editing, which has been widely used in eukaryotic systems, prokaryotic systems, and plants. However, the off-target effects caused by Cas9/sgRNA remain a major concern. Currently, the efforts to reduce the off-target effects mainly focus on improving the targeting specificity of sgRNA/Cas9, regulating the activity of the Cas9 protein or the sgRNA, and controlling the time window of their expression. In this study, a novel system was established to regulate the post-transcriptional sgRNA level by small molecule-controlled aptazyme. This system was shown to reduce the off-target effects caused by Cas9/sgRNA, while enabling precise temporal control over gene editing and regulatory activity. This new system could provide a potentially safer and more powerful tool for genome editing and therapeutic application.

KW - Aptazyme

KW - Cas9

KW - Genome editing

KW - Off-target effect

KW - Theophylline

KW - sgRNA

UR - http://www.scopus.com/inward/record.url?scp=85055910640&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85055910640&partnerID=8YFLogxK

U2 - 10.1016/j.jbiotec.2018.10.011

DO - 10.1016/j.jbiotec.2018.10.011

M3 - Article

C2 - 30391232

AN - SCOPUS:85055910640

SN - 0168-1656

VL - 288

SP - 23

EP - 29

JO - Journal of Biotechnology

JF - Journal of Biotechnology

ER -

Aptazyme-mediated direct modulation of post-transcriptional sgRNA level for conditional genome editing and gene expression

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this