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. 2020 Apr 28:6:27.
doi: 10.1038/s41421-020-0165-z. eCollection 2020.

Efficient generation of mouse models with the prime editing system

Yao Liu #  1   2 , Xiangyang Li #  2 , Siting He #  3   4 , Shuhong Huang  1 , Chao Li  1 , Yulin Chen  1 , Zhen Liu  3 , Xingxu Huang  2   5 , Xiaolong Wang  1
Affiliations

Efficient generation of mouse models with the prime editing system

Yao Liu et al. Cell Discov. .
No abstract available

Keywords: Bioinformatics; Genomic analysis.

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Conflict of interest statement

Conflict of interestThe authors declare that they have no conflict of interest.

Figures

Fig. 1
Fig. 1. PE3-mediated efficient base transversion in vitro and in vivo.
a Schematic diagram of target site at the HOXD13 locus. The PAM sequences and the spacer sequences of pegRNA are underlined in red and black, respectively; the nucleotide substitutions are marked in blue. The corresponding targeted codons are shown on the right of sequence. b Editing efficiency and indel generation by PE3 at the +6 position of HOXD13 using pegRNAs containing 15-nt RT templates and PBS sequences ranging from 10 to 17 nt in N2a cells. Indels (gray column) are plotted for comparison. c PE3-mediated base transversion editing efficiency and indels at the +6 position of HOXD13 in mouse N2a cells as a function of RT template length. Indels (gray line) are plotted for comparison. The sequence below the graph shows the last nucleotide template for synthesis by the pegRNA. d Sanger sequencing chromatograms at the HOXD13 site in N2a cells. The PAM sequence and spacer sequence of pegRNA are underlined in red and black, respectively. Asterisks indicate the desired editing. e Frequencies of nucleotide conversions in mouse embryos. PCR amplicons from the target regions in Hoxd13 were analyzed by targeted deep sequencing. Each dot indicates one individual embryo; embryo id was marked for the embryos with top five nucleotide conversion frequency. f Frequencies of nucleotide conversions in mice. PCR amplicons from the target regions in Hoxd13 were analyzed by targeted deep sequencing. Each dot indicates one individual mouse. g Genotypes of the two Hoxd13 sites in mice by targeted deep sequencing. Alignment of sequences derived from deep sequencing in ten mutant mice. The target site is highlighted in blue. pegRNA spacer sequence are underlined in black. PAM sequences are underlined and marked in red.
See this image and copyright information in PMC

References

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