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. 2014 Feb;6(1):97-9.
doi: 10.1093/jmcb/mjt047. Epub 2014 Jan 8.

Effective gene targeting in rabbits using RNA-guided Cas9 nucleases

Dongshan Yang  1 , Jie Xu, Tianqing Zhu, Jianglin Fan, Liangxue Lai, Jifeng Zhang, Y Eugene Chen
Affiliations

Effective gene targeting in rabbits using RNA-guided Cas9 nucleases

Dongshan Yang et al. J Mol Cell Biol. 2014 Feb.
No abstract available

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Figures

Figure 1
Figure 1
Generation of KO rabbits with RNA-guided Cas9 nucleases. (A) Constructs of Cas9 RNA system used in this study. NLS, nuclear localization signal; bGH-pA, bovine growth hormone poly-A; PAM, protospacer adaptor motif. (B) Representative T7 endonuclease 1 assay results of newborn rabbits. T7 endonuclease 1 recognizes and cuts any mismatched bps in the double-strand DNA. Multiple bands on the gel indicate a successful gene targeting events (+). One single band indicates a WT genotype (−). (C) Sanger sequencing of targeted gene alleles in all founder KO rabbits, including CD36 (n = 11), APOE (n = 10), LDLR (n = 9), and RyR2 (n = 8) KO founders. Protospacer (target) sequence (20 nt) is shown in green on the top row for each gene, followed by 3 nt PAM (shown in red). For each gene, the number of founder animals is shown on the left column. Sequence information of the targeted alleles is shown in the middle column, where dash dots indicate deletions and italic red letters indicate insertions. The sizes of the insertions (+) or deletions (Δ) are shown on the right column. LDLR KO founder #1–5 showed the same Δ15 deletion in one allele; therefore they are consolidated into one row. Wt, wild-type. (D) In vitro gene targeting efficiency of Cas9 constructs. (E) Production efficiency of ApoE, LDLR, CD36, and RyR2 KO rabbits.
See this image and copyright information in PMC

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Publication types