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4 results on '"Shou-Long Deng"'

1. High temperature suppressed SSC self-renewal through S phase cell cycle arrest but not apoptosis

Author

Jia Wang, Wei-Jun Gao, Shou-Long Deng, Xiang Liu, Hua Jia, and Wen-Zhi Ma

Subjects
High temperature, SSCs, Self-renewal, Cell cycle arrest, Apoptosis, Medicine (General), R5-920, Biochemistry, QD415-436
Abstract

Abstract Background High temperature has a very adverse effect on mammalian spermatogenesis and eventually leads to sub- or infertility through either apoptosis or DNA damage. However, the direct effects of heat stress on the development of spermatogonial stem cells (SSCs) are still unknown because SSCs are rare in the testes. Methods In the present study, we first used in vitro-cultured SSCs to study the effect of heat shock treatment on SSC development. Then, we used RNA-Seq analysis to identify new genes or signalling pathways implicated in the heat stress response. Results We found that 45 min of 43 °C heat shock treatment significantly inhibited the proliferation of SSCs 2 h after treatment but did not lead to apoptosis. In total, 17,822 genes were identified by RNA-Seq after SSC heat shock treatment. Among these genes, we found that 200 of them had significantly changed expression, with 173 upregulated and 27 downregulated genes. The number of differentially expressed genes in environmental information processing pathways was 37, which was the largest number. We screened the candidate JAK-STAT signalling pathway on the basis of inhibition of cell cycle progression and found that the JAK-STAT pathway was inhibited after heat shock treatment. The flow cytometry results further confirmed that heat stress caused S phase cycle arrest of SSCs. Conclusion Our results showed that heat shock treatment at 43 °C for 45 min significantly inhibited SSC self-renewal through S phase cell cycle arrest but not apoptosis.

Published
2019
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2. CRISPR/Cas9-mediated genome editing induces gene knockdown by altering the pre-mRNA splicing in mice

Author

Ji-Xin Tang, Dafeng Chen, Shou-Long Deng, Jian Li, Yuanyuan Li, Zheng Fu, Xiu-Xia Wang, Yan Zhang, Su-Ren Chen, and Yi-Xun Liu

Subjects
Biotechnology, TP248.13-248.65
Abstract

Abstract Background Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9 (CRISPR/Cas9) has been wildly used to generate gene knockout models through inducing indels causing frame-shift. However, there are few studies concerning the post-transcript effects caused by CRISPR-mediated genome editing. Results In the present study, we showed that gene knockdown model also could be generated using CRISPR-mediated gene editing by disrupting the boundary of exon and intron in mice (C57BL/6 J). CRISPR induced indel at the boundary of exon and intron (5′ splice site) caused alternative splicing and produced multiple different mRNAs, most of these mRNAs introduced premature termination codon causing down expression of the gene. Conclusions These results showed that alternative splicing mutants were able to generate through CRISPR-mediated genome editing by deleting the boundary of exon and intron causing disruption of 5′ splice site. Although alternative splicing was an unexpected outcome, this finding could be developed as a technology to generate gene knockdown models or to investigate pre-mRNA splicing.

Published
2018
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3. CRISPR/Cas9-mediated genome editing induces gene knockdown by altering the pre-mRNA splicing in mice

Author

Su-Ren Chen, Yi-Xun Liu, Yuan-Yuan Li, Jian Li, Shou-Long Deng, Yan Zhang, Dafeng Chen, Xiu-Xia Wang, Zheng Fu, and Ji-Xin Tang

Subjects
0301 basic medicine, RNA Splicing, lcsh:Biotechnology, Biology, Mice, 03 medical and health sciences, Exon, 0302 clinical medicine, INDEL Mutation, Genome editing, lcsh:TP248.13-248.65, RNA Precursors, Animals, CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats, Gene Editing, Genetics, Gene knockdown, Cas9, Methodology Article, Alternative splicing, Intron, Exons, Introns, Mice, Inbred C57BL, 030104 developmental biology, Gene Knockdown Techniques, RNA splicing, CRISPR-Cas Systems, 030217 neurology & neurosurgery, Biotechnology
Abstract

Background Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9 (CRISPR/Cas9) has been wildly used to generate gene knockout models through inducing indels causing frame-shift. However, there are few studies concerning the post-transcript effects caused by CRISPR-mediated genome editing. Results In the present study, we showed that gene knockdown model also could be generated using CRISPR-mediated gene editing by disrupting the boundary of exon and intron in mice (C57BL/6 J). CRISPR induced indel at the boundary of exon and intron (5′ splice site) caused alternative splicing and produced multiple different mRNAs, most of these mRNAs introduced premature termination codon causing down expression of the gene. Conclusions These results showed that alternative splicing mutants were able to generate through CRISPR-mediated genome editing by deleting the boundary of exon and intron causing disruption of 5′ splice site. Although alternative splicing was an unexpected outcome, this finding could be developed as a technology to generate gene knockdown models or to investigate pre-mRNA splicing. Electronic supplementary material The online version of this article (10.1186/s12896-018-0472-8) contains supplementary material, which is available to authorized users.

Published
2018
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4. Effects of over-expression of TLR2 in transgenic goats on pathogen clearance and role of up-regulation of lysozyme secretion and infiltration of inflammatory cells

Author

Hongtao Zhang, Zhengxing Lian, Maosheng Cui, Hai-Juan He, Yuchang Yao, Yu-Feng Liu, Ning Li, Bao-Lu Zhang, Shou-Long Deng, Juncai Fu, and Kun Yu

Subjects
Lipopolysaccharides, Pam3CSK4, Transgene, Inflammation, Biology, Nitric Oxide, Real-Time Polymerase Chain Reaction, Microbiology, Animals, Genetically Modified, chemistry.chemical_compound, Malondialdehyde, medicine, Animals, TLR2, Secretion, Pathogen, Microinjection, Goat Diseases, lcsh:Veterinary medicine, General Veterinary, Histocytochemistry, Goats, General Medicine, veterinary(all), Toll-Like Receptor 2, In vitro, Up-Regulation, Oxidative Stress, chemistry, Over-expression, Immunology, Leukocytes, Mononuclear, RNA, lcsh:SF600-1100, Female, Muramidase, medicine.symptom, Lysozyme, Research Article
Abstract

Background Toll-like receptor 2 (TLR2) is important to host recognition of invading gram-positive microbes. In goats, these microbes can cause serious mastitis, anthrax, tetanus, and other problems. Transgenic goats constitutively over-expressing TLR2 in many tissues serve as a suitable model for the study of the role of TLR2 over-expression in bacterial clearance. Results Capra hircus TLR2 over-expression vector (p3S-LoxP-TLR2) was used to generate transgenic goats by egg microinjection. The integration efficiency was 8.57%. Real-time PCR and immunohistochemical results confirmed that the goats over-expressing the TLR2 gene (Tg) expressed more TLR2 than wild-type goats (WT). Monocyte-macrophages from the bloodstreams of transgenic goats were stimulated with synthetic bacterial lipoprotein (Pam3CSK4) and by the promotion of interleukin-6 (IL-6) and IL-10 expression in vitro. The oxidative damage was significantly reduced, and lysozyme (LZM) secretion was found to be up-regulated. Ear tissue samples from transgenic goats that had been stimulated with Pam3CSK4 via hypodermic injection showed that transgenic individuals can undergo the inflammation response very quickly. Conclusions Over-expression of TLR2 was found to decrease radical damage to host cells through low-level production of NO and MDA and to promote the clearance of invasive bacteria by up-regulating lysozyme secretion and filtration of inflammatory cells to the infected site.

Published
2012

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