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G3 (Bethesda). 2017 Feb 9;7(2):719-722. doi: 10.1534/g3.116.038091.

Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection.

Author information

1
Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
2
Functional and Chemical Genomics Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104.
3
Embryonic Stem Cell and Transgenic Mouse Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
4
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
5
Zebrafish Core, Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
6
Genomics Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
7
Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892 bpavan@mail.nih.gov burgess@mail.nih.gov.
8
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892 bpavan@mail.nih.gov burgess@mail.nih.gov.

Abstract

Cpf1 has emerged as an alternative to the Cas9 RNA-guided nuclease. Here we show that gene targeting rates in mice using Cpf1 can meet, or even surpass, Cas9 targeting rates (approaching 100% targeting), but require higher concentrations of mRNA and guide. We also demonstrate that coinjecting two guides with close targeting sites can result in synergistic genomic cutting, even if one of the guides has minimal cutting activity.

KEYWORDS:

CRISPR; Cpf1 nuclease; genome editing; mouse

PMID:
28040780
PMCID:
PMC5295614
DOI:
10.1534/g3.116.038091
[Indexed for MEDLINE]
Free PMC Article

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