What question did this study set out to answer?

To engineer a self-processing ribozyme-flanked RNA construct capable of producing functional CRISPR guide RNAs independent of standard promoter constraints.

December 30, 2013Journal of Integrative Plant BiologyOpen Access

Self‐processing of ribozyme‐flanked RNAs into guide RNAs in vitro and in vivo for CRISPR‐mediated genome editing

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Authors

YGYangbin GaoUniversity of California, San Diego YZYunde ZhaoUniversity of California, San Diego

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Implication

Experimental study demonstrates self-cleaving ribozyme-flanked guide RNAs enable versatile CRISPR genome editing in yeast, indicating potential for cell- and tissue-specific targeting.

Key Points

To engineer a self-processing ribozyme-flanked RNA construct capable of producing functional CRISPR guide RNAs independent of standard promoter constraints.
Engineered an artificial gene (RGR) encoding a guide RNA flanked at both termini by self-splicing ribozyme sequences.
Assessed transcript self-cleavage and targeted Cas9-mediated DNA cleavage both in vitro and in vivo using yeast models.
Implemented universal primers to generate in vitro guide RNAs coupled with Cas9 protein to assay gene mutations.
Primary RGR transcripts underwent self-catalyzed cleavage to yield mature, functional guide RNAs free of inhibitory end modifications.
Released guide RNAs successfully directed sequence-specific DNA target cleavage by Cas9 both in vitro and in vivo in yeast.
The system operated independently of restrictive U6 and U3 snRNA promoters, enabling universal transcription and rapid mutation detection.

Cite This Study

Gao et al. (2013) studied this question.

synapsesocial.com/papers/6a01df93897643a80dcb167c https://doi.org/10.1111/jipb.12152

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Implication

Experimental study demonstrates self-cleaving ribozyme-flanked guide RNAs enable versatile CRISPR genome editing in yeast, indicating potential for cell- and tissue-specific targeting.

Self‐processing of ribozyme‐flanked RNAs into guide RNAs in vitro and in vivo for CRISPR‐mediated genome editing

Discussion

Member takes

Implication

Key Points

Cite This Study

Self‐processing of ribozyme‐flanked RNAs into guide RNAs in vitro and in vivo for CRISPR‐mediated genome editing

Discussion

Member takes

Implication

Key Points

Cite This Study

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