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. Author manuscript; available in PMC: 2023 Feb 1.
Published in final edited form as: Adv Drug Deliv Rev. 2021 Dec 20;181:114087. doi: 10.1016/j.addr.2021.114087

Table 1.

Advantages and limitations of CRISPR/Cas9 delivery methods.

Delivery method Advantages Limitations
Viral/Plasmid DNA

  • - Rapid/easy production of plasmid DNA

  • - Persistent Cas9 activity

  • - Possible vector integration into host cell DNA (insertional mutagenesis)

  • - Low frequency of on-target editing

  • - High frequency of off-target editing

  • - High cell toxicity

  • - Difficult production of viral vectors

  • - Reliance on host cell machinery for transcription/translation

  • - Host immune response to foreign DNA sequences and viral proteins

  • - Limited packaging capacities
Cas9 mRNA and sgRNA

  • - Transient Cas9 activity

  • - No possible vector integration into host cell DNA

  • - Various delivery methods

  • - Low frequency of on-target editing

  • - High frequency of off-target editing

  • - High cell toxicity

  • - Difficult production of Cas9 mRNA and sgRNA

  • - Reliance on host cell machinery for translation

  • - Poor stability of mRNA

  • - Inability to be spatially or temporally regulated
Cas9/sgRNA RNP

  • - Rapid Cas9 activity

  • - Transient Cas9 activity

  • - No possible vector integration into host cell DNA

  • - High frequency of on-target editing

  • - Low frequency of off-target editing

  • - Low cell toxicity

  • - Various delivery methods

  • - Difficult production of Cas9 protein and sgRNA

  • - Immunogenicity to Cas9 protein

  • - Inability to be spatially or temporally regulated