3 Reasons to Stop Using In Vitro Transcription (IVT) for CRISPR

Are you still using in vitro transcription (IVT) to make your CRISPR guides?

While it was once the trusted method for making CRISPR guides, in vitro transcription (IVT) is no longer the most effective way to generate gRNA for CRISPR experiments.

Here are 3 Reasons why you should stop using in vitro transcribed gRNA:

Reason 1: Low Purity IVT Yields Inconsistent Editing and Increases Off Target Effects
Reason 2: Variable Efficiency IVT is Less Efficient in Primary and Stem Cells
Reason 3: High Total Cost IVT is Labor Intensive and Not Scalable


CRISPRevolution Editing Efficiency vs. IVT
This chart compares average (solid bars) editing efficiencies and their standard deviation (black lines) between 3 replicates of IVT vs. synthetic sgRNA (modified) on a challenging genomic target in HEK293T cells. Industry partner validated data.


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