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A Single Multiplex crRNA Array for FnCpf1-Mediated Human Genome Editing.

Huihui Sun1, Fanfan Li2, Jie Liu2

  • 1School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, State Key Laboratory and Key Laboratory of Vision Science, Ministry of Health and Zhejiang Provincial Key Laboratory of Ophthalmology and Optometry, Wenzhou, Zhejiang 325027, China.

Molecular Therapy : the Journal of the American Society of Gene Therapy
|June 19, 2018
PubMed
Summary

This study shows that FnCpf1 enables multiplex human genome editing using a single CRISPR RNA array. This advance offers a powerful tool for complex genetic research and potential gene therapies.

Keywords:
FnCpf1crRNAmultiplex genome editing

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Cpf1 is a versatile genome editing tool known for its simplicity and efficiency.
  • FnCpf1 offers advantages in human genome editing due to its flexible protospacer adjacent motif (PAM) sequence.
  • Multiplex genome editing is crucial for studying gene families, networks, and multigenic disorders.

Purpose of the Study:

  • To demonstrate FnCpf1-mediated multiplex genome editing in human cells using a single customized CRISPR RNA (crRNA) array.
  • To show the capability of a single crRNA array for targeting multiple sites within a single gene.
  • To establish FnCpf1 as an effective tool for complex human genome manipulation.

Main Methods:

  • Utilized a single customized CRISPR RNA (crRNA) array for FnCpf1 delivery.
  • Performed simultaneous targeting of multiple genes in human cells.
  • Demonstrated targeting of multiple sites within a single gene using the crRNA array.

Main Results:

  • Successfully achieved multiplex genome editing in human cells with FnCpf1 via a single crRNA array.
  • Confirmed the ability to target multiple sites within one gene simultaneously.
  • Validated the efficiency and flexibility of FnCpf1 for complex genomic applications.

Conclusions:

  • FnCpf1 is a potent tool for multiplex genome editing in human cells.
  • A single customized crRNA array simplifies and enhances multiplex genome editing strategies.
  • FnCpf1 holds significant promise for advancing human genome manipulation and gene therapy.