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Retinal Degenerative Diseases XIX pp 109–114Cite as

Generation of an Avian Myeloblastosis Virus (AMV) Reverse Transcriptase Prime Editor

Part of the Advances in Experimental Medicine and Biology book series (AEMB,volume 1415)

Abstract

Prime editing (PE) is a novel, double-strand break (DSB)-independent gene editing technology that represents an exciting avenue for the treatment of inherited retinal diseases (IRDs). Given the extensive and heterogenous nature of the 280 genes associated with IRDs, genome editing has presented countless complications. However, recent advances in genome editing technologies have identified PE to have tremendous potential, with the capability to ameliorate small deletions and insertions in addition to all twelve possible transition and transversion mutations. The current PE system is based on the fusion of the Streptococcus pyogenes Cas9 (SpCas9) nickase H840A mutant and an optimized Moloney murine leukemia virus (MMLV) reverse-transcriptase (RT) in conjunction with a PE guide RNA (pegRNA). In this study, we developed a prime editor based on the avian myeloblastosis virus (AMV)-RT and showed its applicability for the installation of the PRPH2 c.828+1G>A mutation in HEK293 cells.

Keywords

  • Prime editing
  • Avian myeloblastosis virus (AMV)
  • PRPH2
  • Retinitis pigmentosa (RP)
  • AMV-RT prime editors

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References

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Acknowledgments

SHT and the Jonas Children’s Vision Care and Bernard & Shirlee Brown Glaucoma Laboratory are supported by the National Institutes of Health [P30EY019007, R01EY018213, R01EY024698, R01EY026682, R24EY027285, U01EY030580], National Cancer Institute Core [5P30CA013696], Foundation Fighting Blindness [TA-NMT-0116-0692-COLU], the Research to Prevent Blindness (RPB) Physician-Scientist Award. BLD is a recipient of the Capes PhD scholarship. PMJQ is the current recipient of a Curing Retinal Blindness Foundation (CRBF) grant, a Knights Templar Eye Foundation (KTEF) Career Starter grant, an Uplifting Athletes Young Investigator grant, and a New York Stem Cell Foundation (NYSCF) – Druckenmiller Fellowship.

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Author notes

  1. Authors Bruna Lopes da Costa, Salvatore Caruso and Nicolas D Nolan have equally contributed to this chapter.

Authors and Affiliations

  1. Department of Biomedical Engineering, Columbia University, New York, NY, USA

    Yi-Ting Tsai, Bruna Lopes da Costa, Salvatore Marco Caruso, Nicolas D. Nolan & Stephen H. Tsang

  2. Edward S. Harkness Eye Institute, Department of Ophthalmology, Columbia University Irving Medical Center/New York-Presbyterian Hospital, New York, NY, USA

    Bruna Lopes da Costa, Salvatore Marco Caruso, Nicolas D. Nolan, Sarah R. Levi, Stephen H. Tsang & Peter M. J. Quinn

  3. Jonas Children’s Vision Care, and Bernard & Shirlee Brown Glaucoma Laboratory, Department of Ophthalmology, Columbia University, New York, NY, USA

    Bruna Lopes da Costa, Salvatore Marco Caruso, Nicolas D. Nolan, Sarah R. Levi, Stephen H. Tsang & Peter M. J. Quinn

  4. Columbia Stem Cell Initiative, Columbia University, New York, NY, USA

    Salvatore Marco Caruso, Nicolas D. Nolan & Stephen H. Tsang

  5. Department of Pathology & Cell Biology, Columbia University, New York, NY, USA

    Stephen H. Tsang

  6. Institute of Human Nutrition, Columbia University, New York, NY, USA

    Stephen H. Tsang

Authors
  1. Yi-Ting Tsai
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  2. Bruna Lopes da Costa
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  3. Salvatore Marco Caruso
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  4. Nicolas D. Nolan
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  5. Sarah R. Levi
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  6. Stephen H. Tsang
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  7. Peter M. J. Quinn
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Corresponding author

Correspondence to Peter M. J. Quinn .

Editor information

Editors and Affiliations

  1. Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    John D. Ash

  2. Ocular Genomics InstituteDepartment of OphthalmologyMassachusetts Eye and Ear InfirmaryHarvard Medical School, Boston, MA, USA

    Eric Pierce

  3. Health Sciences Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Robert E. Anderson

  4. Department of Ophthalmology, Duke Medical Center, Durham, NC, USA

    Catherine Bowes Rickman

  5. Department of Ophthalmology, Cleveland Clinic Lerner College of Medicine, Cleveland, OH, USA

    Joe G. Hollyfield

  6. Laboratory for Retinal Cell BiologyDepartment of OphthalmologyUniversity Hospital Zurich, University of Zurich, Schlieren, Switzerland

    Christian Grimm

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© 2023 The Author(s), under exclusive license to Springer Nature Switzerland AG

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Cite this paper

Tsai, YT. et al. (2023). Generation of an Avian Myeloblastosis Virus (AMV) Reverse Transcriptase Prime Editor. In: Ash, J.D., Pierce, E., Anderson, R.E., Bowes Rickman, C., Hollyfield, J.G., Grimm, C. (eds) Retinal Degenerative Diseases XIX. Advances in Experimental Medicine and Biology, vol 1415. Springer, Cham. https://doi.org/10.1007/978-3-031-27681-1_17

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