articleNature CommunicationsFeb 16, 2017GOLD OA

CRISPR/Cpf1-mediated DNA-free plant genome editing

HKHye‐Ran KimSKSang‐Tae KimJRJahee RyuBKBeum‐Chang KangJKJin‐Soo Kim

Institute for Basic Science · Seoul National University

PubMed
Indexed incrossrefdoajpubmed

Abstract

Cpf1, a type V CRISPR effector, recognizes a thymidine-rich protospacer-adjacent motif and induces cohesive double-stranded breaks at the target site guided by a single CRISPR RNA (crRNA). Here we show that Cpf1 can be used as a tool for DNA-free editing of plant genomes. We describe the delivery of recombinant Cpf1 proteins with in vitro transcribed or chemically synthesized target-specific crRNAs into protoplasts isolated from soybean and wild tobacco. Designed crRNAs are unique and do not have similar sequences (≤3 mismatches) in the entire soybean reference genome. Targeted deep sequencing analyses show that mutations are successfully induced in FAD2 paralogues in soybean and AOC in wild tobacco. Unlike…

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Topics & keywords

Topics
Keywords
  • Trans-activating crRNA
  • CRISPR
  • Genome editing
  • Biology
  • Genome
  • Effector
  • DNA
  • Genetics
UN Sustainable Development Goals
  • Good health and well-being
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