Fluorescent Guide RNAs Facilitate Development of Layered Pol II-Driven CRISPR Circuits

David J. Menn, Swechchha Pradhan, Samira Kiani, Xiao Wang

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Efficient clustered regularly interspaced short palindromic repeat (CRISPR) guide RNA (gRNA) expression from RNA Polymerase II (Pol II) promoters will aid in construction of complex CRISPR-based synthetic gene networks. Yet, we require tools to properly visualize gRNA directly to quantitatively study the corresponding network behavior. To address this need, we employed a fluorescent gRNA (fgRNA) to visualize synthetic CRISPR network dynamics without affecting gRNA functionality. We show that studying gRNA dynamics directly enables circuit modification and improvement of network function in Pol II-driven CRISPR circuits. This approach generates information necessary for optimizing the overall function of these networks and provides insight into the hurdles remaining in Pol II-regulated gRNA expression. ©

Original languageEnglish (US)
Pages (from-to)1929-1936
Number of pages8
JournalACS Synthetic Biology
Volume7
Issue number8
DOIs
StatePublished - Aug 17 2018

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Guide RNA
RNA
Networks (circuits)
Synthetic Genes
RNA Polymerase II
Gene Regulatory Networks
Genes

Keywords

  • computational biology
  • CRISPR
  • fgRNA
  • genetic circuit
  • synthetic biology

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

Fluorescent Guide RNAs Facilitate Development of Layered Pol II-Driven CRISPR Circuits. / Menn, David J.; Pradhan, Swechchha; Kiani, Samira; Wang, Xiao.

In: ACS Synthetic Biology, Vol. 7, No. 8, 17.08.2018, p. 1929-1936.

Research output: Contribution to journalArticle

Menn, David J. ; Pradhan, Swechchha ; Kiani, Samira ; Wang, Xiao. / Fluorescent Guide RNAs Facilitate Development of Layered Pol II-Driven CRISPR Circuits. In: ACS Synthetic Biology. 2018 ; Vol. 7, No. 8. pp. 1929-1936.
@article{c63d5c842259497fb25c21bb165b09be,
title = "Fluorescent Guide RNAs Facilitate Development of Layered Pol II-Driven CRISPR Circuits",
abstract = "Efficient clustered regularly interspaced short palindromic repeat (CRISPR) guide RNA (gRNA) expression from RNA Polymerase II (Pol II) promoters will aid in construction of complex CRISPR-based synthetic gene networks. Yet, we require tools to properly visualize gRNA directly to quantitatively study the corresponding network behavior. To address this need, we employed a fluorescent gRNA (fgRNA) to visualize synthetic CRISPR network dynamics without affecting gRNA functionality. We show that studying gRNA dynamics directly enables circuit modification and improvement of network function in Pol II-driven CRISPR circuits. This approach generates information necessary for optimizing the overall function of these networks and provides insight into the hurdles remaining in Pol II-regulated gRNA expression. {\circledC}",
keywords = "computational biology, CRISPR, fgRNA, genetic circuit, synthetic biology",
author = "Menn, {David J.} and Swechchha Pradhan and Samira Kiani and Xiao Wang",
year = "2018",
month = "8",
day = "17",
doi = "10.1021/acssynbio.8b00153",
language = "English (US)",
volume = "7",
pages = "1929--1936",
journal = "ACS Synthetic Biology",
issn = "2161-5063",
publisher = "American Chemical Society",
number = "8",

}

TY - JOUR

T1 - Fluorescent Guide RNAs Facilitate Development of Layered Pol II-Driven CRISPR Circuits

AU - Menn, David J.

AU - Pradhan, Swechchha

AU - Kiani, Samira

AU - Wang, Xiao

PY - 2018/8/17

Y1 - 2018/8/17

N2 - Efficient clustered regularly interspaced short palindromic repeat (CRISPR) guide RNA (gRNA) expression from RNA Polymerase II (Pol II) promoters will aid in construction of complex CRISPR-based synthetic gene networks. Yet, we require tools to properly visualize gRNA directly to quantitatively study the corresponding network behavior. To address this need, we employed a fluorescent gRNA (fgRNA) to visualize synthetic CRISPR network dynamics without affecting gRNA functionality. We show that studying gRNA dynamics directly enables circuit modification and improvement of network function in Pol II-driven CRISPR circuits. This approach generates information necessary for optimizing the overall function of these networks and provides insight into the hurdles remaining in Pol II-regulated gRNA expression. ©

AB - Efficient clustered regularly interspaced short palindromic repeat (CRISPR) guide RNA (gRNA) expression from RNA Polymerase II (Pol II) promoters will aid in construction of complex CRISPR-based synthetic gene networks. Yet, we require tools to properly visualize gRNA directly to quantitatively study the corresponding network behavior. To address this need, we employed a fluorescent gRNA (fgRNA) to visualize synthetic CRISPR network dynamics without affecting gRNA functionality. We show that studying gRNA dynamics directly enables circuit modification and improvement of network function in Pol II-driven CRISPR circuits. This approach generates information necessary for optimizing the overall function of these networks and provides insight into the hurdles remaining in Pol II-regulated gRNA expression. ©

KW - computational biology

KW - CRISPR

KW - fgRNA

KW - genetic circuit

KW - synthetic biology

UR - http://www.scopus.com/inward/record.url?scp=85050334592&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85050334592&partnerID=8YFLogxK

U2 - 10.1021/acssynbio.8b00153

DO - 10.1021/acssynbio.8b00153

M3 - Article

C2 - 30021068

AN - SCOPUS:85050334592

VL - 7

SP - 1929

EP - 1936

JO - ACS Synthetic Biology

JF - ACS Synthetic Biology

SN - 2161-5063

IS - 8

ER -