One-step generation of myostatin gene knockout sheep via the CRISPR/Cas9 system

doi:10.15302/J-FASE-2014007

Front. Agr. Sci. Eng.

2014, Vol. 1

Issue (1) : 2-5 https://doi.org/10.15302/J-FASE-2014007

LETTER

One-step generation of myostatin gene knockout sheep via the CRISPR/Cas9 system

Hongbing HAN¹,Yonghe MA²,Tao WANG²,Ling LIAN¹,Xiuzhi TIAN¹,Rui HU¹,Shoulong DENG³,Kongpan LI²,Feng WANG¹,Ning LI²,Guoshi LIU^1,^*(

),Yaofeng ZHAO^2,^*(

),Zhengxing LIAN^1,^*(

)

1. Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China
2. State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China
3. State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China

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Corresponding Author(s): Guoshi LIU

Issue Date: 22 May 2014

Cite this article:

Hongbing HAN,Yonghe MA,Tao WANG, et al. One-step generation of myostatin gene knockout sheep via the CRISPR/Cas9 system[J]. Front. Agr. Sci. Eng. , 2014, 1(1): 2-5.

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https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2014007
https://academic.hep.com.cn/fase/EN/Y2014/V1/I1/2

Fig.1 Generation of MSTN-knockout sheep and analysis of the mutant alleles. (a) A schematic of the sgRNAs targeting the third exon of the MSTN gene. The PAM motif is shown in green. The target site is underlined, and the 12 bp seed sequence is highlighted in blue. The primers for the PCR analysis are indicated by arrows; (b) Indel mutations in the ear and muscle were detected using the Surveyor assay. Ear and muscle tissues from the control group with a wild-type MSTN gene produced a 556 bp band. Monoallelic mutant sheep produced multiple bands with lengths of 556 bp, 331 bp, and 225 bp; (c) Sequence analysis of the mutations detected in the two lambs. Deletions are indicated by a dashed line, and insertions are shaded in cyan. The numbers following the sequences indicate the specific type of mutation, and the clone numbers are surrounded by brackets

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