Purification and in vivo, cell-free, and in vitro characterization of CRISPR-Cas12a2

Methods in enzymology on CD-ROM/Methods in enzymology, Год журнала: 2025, Номер unknown, С. 143 - 181

Опубликована: Янв. 1, 2025

Язык: Английский

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Trans-cleavage activity of Cas12a effectors can be unleashed by both double-stranded DNA and single-stranded RNA targeting in absence of PAM

International Journal of Biological Macromolecules, Год журнала: 2025, Номер unknown, С. 142992 - 142992

Опубликована: Апрель 1, 2025

Язык: Английский

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On the ever‐growing functional versatility of the CRISPR‐Cas13 system

Microbial Biotechnology, Год журнала: 2024, Номер 17(2)

Опубликована: Фев. 1, 2024

CRISPR-Cas systems evolved in prokaryotes to implement a powerful antiviral immune response as result of sequence-specific targeting by ribonucleoproteins. One such consists an RNA-guided RNA endonuclease, known CRISPR-Cas13. In very recent years, this system is being repurposed different ways order decipher and engineer gene expression programmes. Here, we discuss the functional versatility CRISPR-Cas13 system, which includes ability for silencing, editing, tracking, nucleic acid detection translation regulation. This palette makes relevant tool broad field synthetic biology.

Язык: Английский

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An Orthogonal CRISPR/dCas12a System for RNA Imaging in Live Cells

Analytical Chemistry, Год журнала: 2024, Номер 96(15), С. 5913 - 5921

Опубликована: Апрель 2, 2024

CRISPR/Cas technology has made great progress in the field of live-cell imaging beyond genome editing. However, effective and easy-to-use CRISPR systems for labeling multiple RNAs interest are still needed. Here, we engineered a CRISPR/dCas12a system that enables specific recognition target RNA under guidance PAM-presenting oligonucleotide (PAMmer) to mimic PAM mechanism DNA substrates. We demonstrated feasibility specificity this specifically visualizing endogenous mRNA. By leveraging dCas12a-mediated precursor (pre-crRNA) processing orthogonality dCas12a from different bacteria, further proposed as simple versatile molecular toolkit multiplexed types transcripts live cells with high specificity. This programmable not only broadens toolbox but also facilitates diverse applications manipulation.

Язык: Английский

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Trends in developing one-pot CRISPR diagnostics strategies

Trends in biotechnology, Год журнала: 2024, Номер unknown

Опубликована: Авг. 1, 2024

Язык: Английский

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