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Switching the activity of Cas12a using guide RNA strand displacement circuits

Oesinghaus, Lukas ; Simmel, Friedrich C

Nature communications, 2019-05, Vol.10 (1), p.2092-2092, Article 2092 [Periódico revisado por pares]

England: Nature Publishing Group

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Título:
Switching the activity of Cas12a using guide RNA strand displacement circuits
Autor: Oesinghaus, Lukas ; Simmel, Friedrich C
Assuntos: Acidaminococcus - genetics ; Bacterial Proteins - genetics ; Circuits ; CRISPR ; CRISPR-Associated Proteins - genetics ; CRISPR-Associated Proteins - metabolism ; Deoxyribonuclease ; Deoxyribonucleic acid ; Displacement ; DNA ; E coli ; Escherichia coli - genetics ; Gene Editing - methods ; Gene expression ; Gene Expression Regulation, Bacterial - genetics ; Genetic modification ; Logic circuits ; Plasmids - genetics ; Ribonucleic acid ; RNA ; RNA, Guide, CRISPR-Cas Systems - genetics ; Transcription ; Transformation, Bacterial
É parte de: Nature communications, 2019-05, Vol.10 (1), p.2092-2092, Article 2092
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
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Descrição: The CRISPR effector protein Cas12a has been used for a wide variety of applications such as in vivo gene editing and regulation or in vitro DNA sensing. Here, we add programmability to Cas12a-based DNA processing by combining it with strand displacement-based reaction circuits. We first establish a viable strategy for augmenting Cas12a guide RNAs (gRNAs) at their 5' end and then use such 5' extensions to construct strand displacement gRNAs (SD gRNAs) that can be activated by single-stranded RNA trigger molecules. These SD gRNAs are further engineered to exhibit a digital and orthogonal response to different trigger RNA inputs-including full length mRNAs-and to function as multi-input logic gates. We also demonstrate that SD gRNAs can be designed to work inside bacterial cells. Using such in vivo SD gRNAs and a DNase inactive version of Cas12a (dCas12a), we demonstrate logic gated transcriptional control of gene expression in E. coli.
Editor: England: Nature Publishing Group
Idioma: Inglês

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Switching the activity of Cas12a using guide RNA strand displacement circuits

Oesinghaus, Lukas ; Simmel, Friedrich C

England: Nature Publishing Group

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