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Mutation-specific reporter for the optimization and enrichment of prime editing

I.F. Schene, I.P. Joore, J.H.L. Baijens, S. Shehata, E.F. Ilcken, D.P. Bolhuis, R.C.M. van Rees, S.A. Spelier, P.J. van der Doef, J.M. Beekman, E.E.S. Nieuwenhuis, S.A. Fuchs
doi: https://doi.org/10.1101/2021.05.08.443062
I.F. Schene
1Division of Pediatric Gastroenterology, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
2Department of Metabolic Diseases, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
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I.P. Joore
1Division of Pediatric Gastroenterology, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
2Department of Metabolic Diseases, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
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J.H.L. Baijens
4Utrecht University Graduate School of Life Sciences, Heidelberglaan 8, 3584 CS Utrecht, the Netherlands
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S. Shehata
1Division of Pediatric Gastroenterology, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
2Department of Metabolic Diseases, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
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E.F. Ilcken
2Department of Metabolic Diseases, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
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D.P. Bolhuis
2Department of Metabolic Diseases, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
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R.C.M. van Rees
2Department of Metabolic Diseases, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
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S.A. Spelier
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
5Department of Pediatric Respiratory Medicine, Wilhelmina Children’s Hospital, University Medical Center, Utrecht University, 3584 EA Utrecht, the Netherlands
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P.J. van der Doef
6Department of Pediatric Gastroenterology, University Medical Center Groningen, Hanzeplein 1, 9713 GZ Groningen, the Netherlands
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J.M. Beekman
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
5Department of Pediatric Respiratory Medicine, Wilhelmina Children’s Hospital, University Medical Center, Utrecht University, 3584 EA Utrecht, the Netherlands
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E.E.S. Nieuwenhuis
1Division of Pediatric Gastroenterology, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
7Department of Sciences, University College Roosevelt, Lange Noordstraat 1, 4331 CB Middelburg, the Netherlands
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S.A. Fuchs
2Department of Metabolic Diseases, Wilhelmina Children’s Hospital, University Medical Center Utrecht, Lundlaan 6, 3584 EA Utrecht, the Netherlands
3Regenerative Medicine Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands
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  • For correspondence: s.fuchs@umcutrecht.nl
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Abstract

We present a fluorescent prime editing and enrichment reporter (fluoPEER), which can be tailored to any genomic target site. This system rapidly and faithfully ranks the efficiency of prime edit guide RNAs (pegRNAs) and any prime editor protein, including novel variants with flexible PAM recognition. Successful reporter editing enriches for genomic editing. FluoPEER can be employed for efficient correction of patient cells and to elucidate cellular mechanisms needed for successful prime editing.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • ↵a :Lead contact, on behalf of United for Metabolic Diseases (UMD)

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted May 08, 2021.
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Mutation-specific reporter for the optimization and enrichment of prime editing
I.F. Schene, I.P. Joore, J.H.L. Baijens, S. Shehata, E.F. Ilcken, D.P. Bolhuis, R.C.M. van Rees, S.A. Spelier, P.J. van der Doef, J.M. Beekman, E.E.S. Nieuwenhuis, S.A. Fuchs
bioRxiv 2021.05.08.443062; doi: https://doi.org/10.1101/2021.05.08.443062
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Mutation-specific reporter for the optimization and enrichment of prime editing
I.F. Schene, I.P. Joore, J.H.L. Baijens, S. Shehata, E.F. Ilcken, D.P. Bolhuis, R.C.M. van Rees, S.A. Spelier, P.J. van der Doef, J.M. Beekman, E.E.S. Nieuwenhuis, S.A. Fuchs
bioRxiv 2021.05.08.443062; doi: https://doi.org/10.1101/2021.05.08.443062

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