Reproducibility of Indel Formation Rates by Comparing GuideRNA Format and Delivery Method

Multiple configurations of guideRNA and Cas9 components can be used for editing cells. A few options include: a plasmid expressing both the guideRNA and Cas9, Cas9 protein combined with a synthetic single guideRNA, and Cas9 combined with a synthetic 2-part guideRNA. In addition, delivering the compo...

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Bibliographic Details
Published in:Journal of biomolecular techniques 2019-12, Vol.30 (Suppl), p.S34-S35
Main Authors: Delventhal, Kym, Miller, Brandon, Pruett-Miller, Shondra M, Modarai, Shirin, Porter, Shaina N, Regan, Maureen R, Sergison, Elizabeth
Format: Article
Language:English
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Poster Abstracts
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Summary:Multiple configurations of guideRNA and Cas9 components can be used for editing cells. A few options include: a plasmid expressing both the guideRNA and Cas9, Cas9 protein combined with a synthetic single guideRNA, and Cas9 combined with a synthetic 2-part guideRNA. In addition, delivering the components to cells can be done using lipofection or nucleofection transfection methods. In the GERG 2017 survey (https://abrf.org/sites/default/files/gerg_survey_poster_2018_final_0.pdf), plasmid format and lipofection delivery were favored among cell culture users. Meanwhile, RNP format for the guideRNA and Cas9 is gaining in popularity in combination with nucleofection delivery. This study aims to compare cutting efficiency at 3 different guideRNA targets based on the guideRNA format and delivery method across multiple labs. Determining which method or format is the most reproducible will be beneficial. Core facilities or research labs getting started with genome editing could use these results as a benchmark for optimizing their own protocols.
ISSN:1524-0215
1943-4731