Regulating CRISPR/Cas9 Function through Conditional Guide RNA Control

Brown, Wes; Zhou, Wenyuan; Deiters, Alexander

doi:10.1002/cbic.202000423

Cited by 31 publications

(20 citation statements)

References 125 publications

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“…Inducible CRISPR/Cas9 systems have been developed to improve the spatiotemporal resolution of DNA cleavage [ 5 , 6 ]. For controlling the gene-editing system, two main approaches were developed.…”

Section: Introductionmentioning

confidence: 99%

Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles

Semikolenova

Sakovina

Akhmetova

et al. 2021

IJMS

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Here, we proposed a new approach to engineering a photoactivatable CRISPR/Cas9 gene-editing system. The novel nanoCRISPR/Cas9 system is based on the use of auxiliary photocleavable oligodeoxyribonucleotides (PC-DNAs) complementary to crRNA. PC-DNAs contained up to three UV-sensitive linkers made of 1-(2-nitrophenyl)-1,2-ethanediol inside the oligonucleotide chain. Immobilizing PC-DNAs on the surface of carbon nanoparticles through 3′-terminal pyrene residue provided sufficient blocking of crRNA (and corresponding Cas9 activity) before UV irradiation and allows for crRNA release after UV irradiation at 365 nm, which restores Cas9 activity. We optimized the length of blocking photocleavable oligonucleotide, number of linkers, time of irradiation, and the type of carbon nanoparticles. Based on the results, we consider the nanoCRISPR/Cas9 system involving carbon-encapsulated iron nanoparticles the most promising. It provides the greatest difference of functional activity before/after irradiation and can be used in prospective for magnetic field-controlled delivery of CRISPR system into the target cells or tissues and spatiotemporal gene editing induced by UV irradiation.

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Section: Introductionmentioning

confidence: 99%

Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles

Semikolenova

Sakovina

Akhmetova

et al. 2021

IJMS

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“…In this concept, we focus on the recent progresses in inducible CRISPR‐dCas9 transcriptional systems and summarize the approaches that have been developed to control the activities and functions of gRNAs or dCas9 proteins. For readers who are interested in regulating the activities of the CRISPR‐Cas9 systems, there were several comprehensive reviews [12–13,18–20] …”

Section: Introductionmentioning

confidence: 99%

“…For readers who are interested in regulating the activities of the CRISPR-Cas9 systems, there were several comprehensive reviews. [12][13][18][19][20]…”

Section: Introductionmentioning

confidence: 99%

“…For controlling gRNA function, a general approach is to develop switchable gRNA structures that can restore active conformations in the presence of small-molecule ligands, light, synthetic and endogenous nucleic acids, and nuclease. [12] For controlling the function of dCas9 protein, the recruitment of transcriptional effectors and the complementation of split protein fragments can also be triggered by light, small-molecule ligands and endogenous nucleic acids. [13] A variety of transcriptional reporters have been exploited as the signal outputs, including fluorescent proteins, luciferase, endogenous genes and even light-up RNAs.…”

Section: Introductionmentioning

confidence: 99%

“…Current strategies mainly rely on dynamic engineering of dCas9 protein and gRNA. For controlling gRNA function, a general approach is to develop switchable gRNA structures that can restore active conformations in the presence of small‐molecule ligands, light, synthetic and endogenous nucleic acids, and nuclease [12] . For controlling the function of dCas9 protein, the recruitment of transcriptional effectors and the complementation of split protein fragments can also be triggered by light, small‐molecule ligands and endogenous nucleic acids [13] .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Inducible CRISPR‐dCas9 Transcriptional Systems for Sensing and Genome Regulation

Wang

Jiang

2021

ChemBioChem

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The clustered, regularly interspaced short palindromic repeats‐associated protein 9 endonuclease (CRISPR‐Cas9) and the nuclease‐deactivated Cas9 (dCas9) systems have revolutionized our ability to precisely engineer and regulate genomes. Inducible CRISPR‐dCas9‐based transcriptional systems have been rapidly developed to conditionally control genetic manipulation. Current strategies mainly focus on conditional control of gRNA function and dCas9 protein using exogenous and endogenous triggers, including external light, small molecules, synthetic and intracellular oligonucleotides. These strategies have established novel platforms for the spatiotemporal regulation of genome activation and repression, epigenome editing, and so on. Herein, we summarize the recent progress in conditionally controlling CRISPR‐dCas9 transcriptional systems through gRNA modulation and dCas9 protein engineering.

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START: A Versatile Platform for Bacterial Ligand Sensing with Programmable Performances

Kim,

Seo,

Lim

et al. 2024

Advanced Science

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Recognition of signaling molecules for coordinated regulation of target genes is a fundamental process for biological systems. Cells often rely on transcription factors to accomplish these intricate tasks, yet the subtle conformational changes of protein structures, coupled with the complexity of intertwined protein interaction networks, pose challenges for repurposing these for bioengineering applications. This study introduces a novel platform for ligand‐responsive gene regulation, termed START (Synthetic Trans‐Acting Riboswitch with Triggering RNA). Inspired by the bacterial ligand sensing system, riboswitch, and the synthetic gene regulator, toehold switch, the START platform enables the implementation of synthetic biosensors for various ligands. Rational sequence design with targeted domain optimization yields high‐performance STARTs with a dynamic range up to 67.29‐fold and a tunable ligand sensitivity, providing a simple and intuitive strategy for sensor engineering. The START platform also exhibits modularity and composability to allow flexible genetic circuit construction, enabling seamless implementation of OR, AND, and NOT Boolean logic gates for multiple ligand inputs. The START design principle is capable of broadening the suite of synthetic biosensors for diverse chemical and protein ligands, providing a novel riboregulator chassis for synthetic biology and bioengineering applications.

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Regulating CRISPR/Cas9 Function through Conditional Guide RNA Control

Cited by 31 publications

References 125 publications

Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles

Photoactivatable nanoCRISPR/Cas9 System Based on crRNA Reversibly Immobilized on Carbon Nanoparticles

Inducible CRISPR‐dCas9 Transcriptional Systems for Sensing and Genome Regulation

START: A Versatile Platform for Bacterial Ligand Sensing with Programmable Performances

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