Technology

Arrayed CRISPR and RNAi screening perfected

Scientist looking at imagin results

Imagine you could perform thousands of gene editing experiments on your lab bench in one afternoon. This is all possible with Persomics’ technology.

Persomics ImagineArrays™ enable researchers to edit or silence thousands of genes on a single plate. The ImagineArray™ is a ready-to-run platform with pre-printed experiments that can simultaneously edit or silence up to 3200 genes, which makes it ideal for straight-forward screening as well as advanced cell biology assays such as drug/gene interaction screens and synthetic lethality.

The Persomics printing technology is reagent-agnostic and a wide range of reagents can be printed. Contact us for custom prints!

Our current product range includes:

CRISPR

CRISPR

siRNA

siRNA

miRNA

mirNA

Revolutionary transfection system

Phenotypic screening is an important research tool and underlies most first-in-class drugs. RNAi screening remains one of the most widely used technologies for conducting target identification and validation, and is lately complemented by CRISPR, one of the most powerful technologies for cell biology invented in recent years. Conventional screens are based on microtiter plates and depend on the use of robots, but cost has been a major limiting factor in scaling its use.

On the Persomics ImagineArray™ platform, microtiter wells are replaced with contact-printed spots. Each spot is a unique experiment that edits or silences a single gene in cells growing above the RNA spot. Each spot encapsulates an individual or pooled gRNA, siRNA or miRNA and all reagents needed for transfection.

ImagineArray™Removing the need for physical barriers between individual experiments opens the door to miniaturizing the screening platform. Persomics prints circular spots with a diameter of 350 μm that are placed approximately 500 μm apart. Even at such small distances there is no measurable cross-over between spots. The individual RNA in each spot transfects only the cells growing immediately above it.

Miniaturizing each individual experiment allows us to print whole libraries on a single plate, for instance gRNA/siRNA targeting all kinases in the human genome are printed on approximately 2.06 cm2 (0.32 sq. in.).

Drastically reducing the scale of screens allows researchers to dramatically increase the scale of screening.

Reverse Transfection

Reverse transfection – the method whereby 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 of the amount of reagents required in well-based screening, and the time required per experiment is drastically reduced because experiments are performed simultaneously.

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

CRISPR and RNAi have shown great potential in reverse transfection and based on the methods from Whitehead Institute and Sabatini[1], Persomics provides accessible and fast screening tools.

The Persomics platform has been optimized for gRNAs/siRNAs from several leading providers and a range of transfection reagents. With Persomics ImagineArray™, scientists can

  • edit or silence up to 3200 genes in one single plate
  • run a screen in as little as 48-72h (longer time course also possible)
  • analyze using any fluorescence microscope or standard imaging system

 


More and better data - from cell death to visible phenotypes

Today CRISPR screens are typical done pooled with complex deconvolution focusing on cell death only. With arrayed screening using Persomics ImagineArrayTM plates and high-resolution imaging, the readout generates rich data and the possibility to also analyze all phenotypes by morphology. 

Persomics enables

  • Rich data beyond level of cell death
  • Massive number of experiments performed per plate
  • Less data heterogeneity because cells are under consistent assay conditions across the entire plate
  • Higher data quality because you can run more replicates
  • Wider data sets because you can perform multiple screens under different conditions
  • Straightforward analysis using commercially or freely available analysis tools, such as CellProfiler.

No robotics or liquid handling needed

Today, performing high-throughput CRISPR and RNAi screens requires investments in expertise, time, automation hardware, reagents, plates and other consumables. These investments present barriers for individual research labs to begin using CRISPR or RNAi screens as a research tool, and for existing users to increase use of CRISPR and RNAi screening in their research. With ImagineArray™ these barriers are eliminated.

Using high-density reverse-transfection arrays, gene-editing/silencing screens with ImagineArray™ are faster due to the high number of experiments performed in parallel, while simultaneously using less consumables. Each spot contains all that is needed to run an individual CRISPR or RNAi experiment. Since there are no physical barriers between experiments, cells see the same assay conditions across the entire plate, giving less data heterogeneity. By streamlining the screening process, entire libraries can be screened many times using identical or multiple conditions to generate large data sets.

ImagineArray™ does not require extra equipment beyond what is already present in most laboratories. Designed from the ground up to be optimal for imaging, ImagineArray™ conforms to the SBS microplate specification so they can be used with high content imaging systems and all standard microscopes.

Persomics Printing Technology

In initial adoptions of reverse transfection, arrays were printed iteratively, using DNA spotting technology. The process was time consuming and wasteful of sample.

Scientist printing ImagineArray™

Persomics printing technology is based on a moveable-type approach. Pre-assembled types are stacked to form a single print head of capillaries that prints an entire array in one contact with the substrate. Through this massively parallel printing method we can print 3200 experiments in less than 2.5 seconds, and print 1000 ImagineArrays™ in a single run.

Persomics technology dispenses reagents accurately and reproducibly even when handling difficult liquids. Since each spot of the array has its own capillary, entire libraries can be printed without wasting valuable material. Each capillary is disposable and has its own reagent reservoir so there is no cross-contamination of reagents between adjacent capillaries or between printing runs.

Printed arrays are desiccated and can be stored for up to 14 months without any decrease in quality or silencing performance.

Endless possibilities

With our flexible printing technology there is no limit to the complexity of printed arrays. Our method enables us to print pre-configured experiments as well as quickly respond to client requests for custom prints.

Persomics is continuing to develop and expand the capabilities of the ImagineArray™ platform. We are extending the platform to include CRISPR, developing substrates that concentrate cells on spots, and expanding into combinatorial applications. Even with more than 10 years of research and development, we are still only at the beginning.

 

ImagineArray™ Essentials

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