Search Results

You are looking at 1 - 2 of 2 items for

  • Author: Jianguo Zhao x
  • Refine by access: All content x
Clear All Modify Search
Dengfeng Bi School of Life Sciences, University of Science and Technology of China, Hefei, China
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China

Search for other papers by Dengfeng Bi in
Google Scholar
PubMed
Close
,
Jing Yao State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China
Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, China
University of Chinese Academy of Sciences, Beijing, China

Search for other papers by Jing Yao in
Google Scholar
PubMed
Close
,
Yu Wang State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China
University of Chinese Academy of Sciences, Beijing, China

Search for other papers by Yu Wang in
Google Scholar
PubMed
Close
,
Guosong Qin State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China

Search for other papers by Guosong Qin in
Google Scholar
PubMed
Close
,
Yunting Zhang State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China

Search for other papers by Yunting Zhang in
Google Scholar
PubMed
Close
,
Yanfang Wang Laboratory of Animal (Poultry) Genetics Breeding and Reproduction, Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China

Search for other papers by Yanfang Wang in
Google Scholar
PubMed
Close
, and
Jianguo Zhao School of Life Sciences, University of Science and Technology of China, Hefei, China
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China
Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, China
University of Chinese Academy of Sciences, Beijing, China

Search for other papers by Jianguo Zhao in
Google Scholar
PubMed
Close

An efficient mRNA knockdown strategy is needed to explore gene function in cells and embryos, especially to understand the process of maternal mRNA decay during early embryo development. Cas13, a novel RNA-targeting CRISPR effector protein, could bind and cleave complementary single-strand RNA, which has been employed for mRNA knockdown in mouse and human cells and RNA-virus interference in plants. Cas13 has not yet been reported to be used in pigs. In the current study, we explored the feasibility of CRISPR/Cas13d-mediated endogenous RNA knockdown in pigs. KDM5B, a histone demethylase of H3K4me3, was downregulated at the transcriptional level by 50% with CRISPR/Cas13d in porcine fibroblast cells. Knockdown of KDM5B-induced H3K4me3 expression and decreased the abundance of H3K27me3, H3K9me3, H3K4ac, H4K8ac, and H4K12ac. These changes affected cell proliferation and cell cycle. Furthermore, stable integration of the CRISPR/Cas13d system into the porcine genome resulted in the continuous expression of Cas13d and persistent knockdown of KDM5B. Finally, the RNA-targeting potential of Cas13d was further validated in porcine parthenogenetic embryos. By microinjection of Cas13d mRNA and gRNA targeting KDM5B into porcine oocytes, the expression of KDM5B was downregulated, the abundance of H3K4me3 increased as expected, and the expression of embryonic development-related genes was changed accordingly. These results indicate that CRISPR/Cas13d provides an easily programmable platform for spatiotemporal transcriptional manipulation in pigs.

Open access
Jiaojiao Huang State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China
State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Jiaojiao Huang in
Google Scholar
PubMed
Close
,
Hongyong Zhang State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China
State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Hongyong Zhang in
Google Scholar
PubMed
Close
,
Jing Yao State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Jing Yao in
Google Scholar
PubMed
Close
,
Guosong Qin State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Guosong Qin in
Google Scholar
PubMed
Close
,
Feng Wang State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Feng Wang in
Google Scholar
PubMed
Close
,
Xianlong Wang State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Xianlong Wang in
Google Scholar
PubMed
Close
,
Ailing Luo State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Ailing Luo in
Google Scholar
PubMed
Close
,
Qiantao Zheng State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China
State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Qiantao Zheng in
Google Scholar
PubMed
Close
,
Chunwei Cao State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China
State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Chunwei Cao in
Google Scholar
PubMed
Close
, and
Jianguo Zhao State Key Laboratory of Stem Cell and Reproductive Biology, University of Chinese Academy of Sciences, College of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China

Search for other papers by Jianguo Zhao in
Google Scholar
PubMed
Close

Accumulating evidence suggests that faulty epigenetic reprogramming leads to the abnormal development of cloned embryos and results in the low success rates observed in all mammals produced through somatic cell nuclear transfer (SCNT). The aberrant methylation status of H3K9me and H3K9me2 has been reported in cloned mouse embryos. To explore the role of H3K9me2 and H3K9me in the porcine somatic cell nuclear reprogramming, BIX-01294, known as a specific inhibitor of G9A (histone-lysine methyltransferase of H3K9), was used to treat the nuclear-transferred (NT) oocytes for 14–16 h after activation. The results showed that the developmental competence of porcine SCNT embryos was significantly enhanced both in vitro (blastocyst rate 16.4% vs 23.2%, P<0.05) and in vivo (cloning rate 1.59% vs 2.96%) after 50 nm BIX-01294 treatment. BIX-01294 treatment significantly decreased the levels of H3K9me2 and H3K9me at the 2- and 4-cell stages, which are associated with embryo genetic activation, and increased the transcriptional expression of the pluripotency genes SOX2, NANOG and OCT4 in cloned blastocysts. Furthermore, the histone acetylation levels of H3K9, H4K8 and H4K12 in cloned embryos were decreased after BIX-01294 treatment. However, co-treatment of activated NT oocytes with BIX-01294 and Scriptaid rescued donor nuclear chromatin from decreased histone acetylation of H4K8 that resulted from exposure to BIX-01294 only and consequently improved the preimplantation development of SCNT embryos (blastocyst formation rates of 23.7% vs 21.5%). These results indicated that treatment with BIX-01294 enhanced the developmental competence of porcine SCNT embryos through improvements in epigenetic reprogramming and gene expression.

Free access