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Alone out there? Printing gRNA/CRISPR in reverse transfection

Posted by Martin Svensson, Mar 13, 2017

Reverse transfection – the method where nucleic acids are localized on a substrate prior to addition of cells – was first developed in 2001 at Whitehead Institute for Biomedical Research by Junald Ziauddin and David M. Sabatini[1].

Early adaptations of the initial technology were used in functional genomics screens identifying host genes affecting HIV[2] and Trypanosome infection[3].

Reverse transfection offers multiple benefits of scale. Each experiment requires only a fraction amount of reagents required in well-based screening, and the time required per is experiment is drastically reduced because experiments are performed simultaneously.

A major barrier of adopting this method has been the lack of accurate and efficient printing technology. But, with ImagineArray™, Persomics removes this barrier.

Fall 2016 Persomics started to explore the feasibility and opportunity of performing CRISPR based experiment in printed spots using its unique reverse transfection arrays. The result until today have shown great promise and several of Persomics collaboration partners have started to experience the benefit of arrayed CRISPR screening.

As we are writing this update Persomics is collaborating with large Pharma, leading cell biology institutes as well as leading tools and reagent companies, all interested in the opportunity of making CRISPR screening more accessible. Persomics technology allows scientist to pick a pre-printed plate and quickly screen for a gene/drug interaction, a few cell lines or another application of interest.

Editing on the Persomics ImagineArray™ has proven to work exceptionally well with synthetic gRNA. At medium conditions in engineered HeLa Cas9 cells, we see editing of above 75 % after 96-120 hours, and typically around 40-50 % after 72 hours.

 

CRISPR on ImagineArray™

To our knowledge the results below are among the first CRISPR editing experiment done at scale in dried printed reverse transfection spots. As we are certain this method has a great future we are keen to get in touch with other research groups interested in experiencing the tools and possibly contribute to further development.

Relevant further reading

 


 

References

Topics: Science

Martin Svensson

Written by Martin Svensson

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