李劲松--中国科学院分子细胞科学卓越创新中心

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李劲松
研究员，中科院院士，博士生导师，细胞生物学国家重点实验室主任
E-mail: jsli@@sibcb.ac.cn
实验室主页:

个人简介：

　　1993年7月毕业于江西农业大学畜牧兽医系，获理学学士学位；1996年7月毕业于扬州大学畜牧兽医学院，获理学硕士学位；2002年7月毕业于中国科学院动物研究所，获理学博士学位;2002年8月至2007年7月在美国洛克菲勒大学从事博士后研究；2007年8月起任中科院上海生科院生物化学与细胞生物学研究所研究员，研究组长。2012年获“杰出青年基金”支持。

社会任职：

研究方向：

干细胞与胚胎发育

研究工作：

　　生殖细胞不能在体外培养和增殖，极大地限制了生殖发育生物学发展。2012年，实验室利用核移植和干细胞技术，成功地建立了可无限增殖的小鼠孤雄单倍体胚胎干细胞系(又称为类精子干细胞)，并证明其可功能性地代替精子繁育健康小鼠，创建了半克隆技术;2015年，我们发现H19-DMR和IG-DMR雄性印记基因表达调控区域是半克隆胚胎发育的主要障碍，通过敲除这两个区域使健康半克隆小鼠出生率提高了10倍;实验室还建立了人的类精子干细胞及食蟹猴和人孤雌单倍体胚胎干细胞。这些系统性研究成果产生了集单倍体性、自我更新、多能性和“受精”能力为一体的新型干细胞，为揭示有性繁殖的奥秘、深入进行生殖发育的转化研究和造福人类生殖健康的重要工具。
　　基因编辑是研究其功能的主要手段，然而，哺乳动物的精准复杂遗传改造仍然困难重重。类精子干细胞突破了精子不能被遗传编辑的瓶颈，可携带不同的基因修饰或基因编辑器，并通过“授精”传递到小鼠胚胎中，为精准和复杂基因编辑小鼠的构建及在体遗传筛选奠定了基础。2015年以来，我们利用用该技术体系，围绕发育生殖、疾病模拟、染色体工程等开展应用研究，取得了一系列成果，拓展了哺乳动物遗传改造研究的领域，包括：
　　1. 结合类精子干细胞与单碱基编辑系统，实现了原始生殖细胞发育必需蛋白质DND1关键氨基酸的靶向突变筛选，建立了在体关键氨基酸的遗传筛选方法。
　　2. 结合类精子干细胞和CRISPR-Cas9基因编辑文库，实现了骨骼发育关键基因的靶向遗传筛选。
　　3. 在类精子干细胞中敲除多个基因，通过“授精”一步获得了多基因杂合敲除小鼠，模拟了人类复杂疾病Ⅰ型强直性肌营养不良症(DM1)的大部分病症，证实多基因剂量不足是DM1关键病因，为快速构建复杂疾病小鼠模型提供了新策略。
　　4. 在类精子干细胞上开展染色体改造，获得多个携带19对染色体的小鼠模型，模拟了自然界中最常见的染色体演化事件，为在哺乳动物中开展染色体改造研究奠定了基础。
　　5. 全基因组蛋白质标签小鼠资源库将极大地促进生命科学研究，但传统方法难以实现该资源库构建。我们启动了基因组标签计划(genome tagging project, GTP)，利用类精子干细胞进行基因原位打靶，建立全蛋白质组标签细胞系，通过“授精”可获得蛋白质的标签小鼠，有望为促进生物医学研究提供有价值的资源。
　　目前，实验室围绕单倍体胚胎干细胞及半克隆技术开展四方面研究：
　　1. 单倍体胚胎干细胞自身带来的问题，如自发二倍体化的机制等;
　　2. 类精子干细胞作为精子替代物的问题，如，如何进一步提高半克隆胚胎的发育潜力等;
　　3. 半克隆技术在构建复杂遗传改造小鼠模型中的应用;
　　4. 生殖细胞发育的调控网络研究。

承担科研项目情况：

代表论著：

（#Co-first authors, *: Correspondence authors）
- 一、类精子干细胞介导半克隆技术相关论文（2012年—至今）
1. Zhang XM^#, Yan M^#, Yang Z^#, Xiang H^#, Tang W, Cai X, Wu Q, Liu X, Pei G & Li J*. Creation of artificial karyotypes in mice reveals robustness of genome organization. Cell Res 32(11)(2022), 1026–1029.
2. Zhang H^#, Li Y^#, Ma Y^#, Lai C, Yu Q, Shi G, Li J*. Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells. Protein & Cell 13(2)(2021), 102-119.
3. Zhang XM^#, Wu K^#, Zheng Y^#, Zhao H^#, Gao J, Hou Z, Zhang M, Liao J, Zhang J, Gao Y, Li Y, Li L, Tang F*, Chen ZJ* & Li J*. In vitro expansion of human sperm through nuclear transfer. Cell Research 30(4)(2020), 356-359.
4. Yin Q^#, Wang H^#, Li N^#, Ding Y^#, Xie Z^#, Jin, L^#, Li Y, Wang Q, Liu X, Xu L, Li Q, Ma Y, Cheng Y, Wang K, Zhong C, Yu Q, Tang W, Chen W, Yang W, Zhang F, Ding C, Bao L, Zhou B, Hu P* & Li J*. Dosage effect of multiple genes accounts for multisystem disorder of myotonic dystrophy type 1. Cell Research 30(2)(2020), 133-145. Cover story
5. Li Q^#, Li Y^#, Yin Q^#, Huang S^#, Wang K^#, Zhuo L, Li W, Chang B & Li J*. Temporal regulation of prenatal embryonic development by paternal imprinted loci. Sci China Life Sci 63(1)(2020), 1-17. Cover story
6. Wang L^#, Zhang Y^#, Fu X^#, Dong S^#, Tang S, Zhang N, Song C, Yang N, Zhang L, Wang H, Shi H, Jin L, Zhang F*, Li J* & Hua K*. Joint utilization of genetic analysis and semi-cloning technology reveals a digenic etiology of Mu？llerian anomalies. Cell Res 30(1)(2020), 91-94.
7. Bai M^#, Han Y^#, Wu Y, Liao J, Li L, Wang L, Li Q, Xing w, Chen L, Zou W* & Li J*. Targeted genetic screening in mice through haploid embryonic stem cells identifies critical genes in bone development. PLOS Biology 17(7)(2019), e3000350.
8. Jiang J, Yan M, Li D & Li J*. Genome tagging project: tag every protein in mouse through “artificial spermatids”. Nat Sci Rev 6(3)(2019), 394-396.
9. Li Q^#, Li Y^#, Yang S, Huang S, Yan M, Ding Y, Tang W, Lou X, Yin Q, Sun Z, Lu L, Shi H, Wang H, Chen Y* & Li J*. CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 amino acids that are critical for primordial germ cell development. Nat Cell Biol 20(11)(2018), 1315-1325.
10. Qu C, Yan M, Yang S, Wang L, Yin Q, Liu Y, Chen Y & Li J*. Haploid embryonic stem cells can be enriched and maintained by simple filtration. J Biol Chem 293 (14) (2018), 5230-5235.
11. Wei L, Wang X, Yang S, Yuan W* & Li J*. Efficient generation of the mouse model with a defined point mutation through haploid cell-mediated gene-editing. J Genet Genomics 44(9)(2017), 461-463.
12. Xu H, Yue C, Zhang T, Li Y, Guo A, Liao J, Pei G, Li J* & Jing N*. Derivation of haploid neurons from mouse androgenetic haploid embryonic stem cells. Neurosci Bull 33(3)(2017), 361-364.
13. Zhong C ^#, Zhang M^#, Yin Q, Zhao H, Wang Y, Huang S, Tao W, Wu K*, Chen Z-J* & Li J*. Generation of human haploid embryonic stem cells from parthenogenetic embryos obtained by microsurgical removal of male pronucleus. Cell Res 26(6)(2016), 743-746.
14. Zhong C ^#, Xie Z^#, Yin Q^#, Dong R, Yang S, Wu Y & Li J*. Parthenogenetic haploid embryonic stem cells efficiently support mouse generation by oocyte injection. Cell Res 26(1)(2016), 131-134.
15. Zhong C ^#, Yin Q^#, Xie Z^#, Bai M^#, Dong R^#, Tang W, Xing Y, Zhang H, Yang S, Chen L-L, Bartolomei MS, Ferguson-Smith A, Li D, Yang L*, Wu Y* & Li J*. CRISPR-Cas9-mediated genetic screening in mice with haploid embryonic stem cells carrying a guide RNA library. Cell Stem Cell 17(2)(2015), 221-232.
16. Yang H^#, Liu Z^#, Ma Y^#, Zhong C, Yin Q, Zhou C, Shi L, Cai Y, Zhao H, Wang H, Tang F, Wang Y, Zhang C, Liu X, Lai D, Jin Y*, Sun Q* & Li J*. Generation of haploid embryonic stem cells from Macaca fascicularis monkey parthenotes. Cell Res 23(10)(2013), 1187-1200.
17. Yang H^#, Shi L^#, Wang B^#, Liang D, Zhong C, Liu W, Nie Y, Liu J, Zhao J, Gao X, Li D, Xu G-L* & Li J*. Generation of genetically modified mice by oocyte injection of androgenetic haploid embryonic stem cells. Cell 149(3)(2012), 605-617.
  - 二、生殖细胞发育、基因编辑及治疗相关论文（2013年—至今）
18. Li Q^#, Lu J^#, Yin X^#, Chang Y^#, Wang C^#, Yan M^#, Feng L, Cheng Y, Gao Y, Xu B, Zhang Y, Wang Y, Cui G, Xu L, Sun Y, Zeng R, Li Y, Jing N, Xu GL*, Wu L*, Tang F* & Li J*. Base editing-mediated one-step inactivation of the Dnmt gene family reveals critical roles of DNA methylation during mouse gastrulation. Nat Commun 14(1)(2023): 2922.
19. Fu J^#, Li Q^#, Liu X^#, Tu T^#, Lv X, Yin X, Lv J, Song Z, Qu J, Zhang J, Li J* & Gu F*. Human cell based directed evolution of adenine base editors with improved efficiency. Nat Commun 12(1)(2021), 5897.
20. Zhao T^#, Li Q^#, Zhou C^#, Lv X^#, Liu H, Tu T, Tang N, Cheng Y, Liu X, Liu C, Zhao J, Song Z, Wang H, Li J* & Gu F*. Small-molecule compounds boost genome-editing efficiency of cytosine base editor. Nucleic Acids Res 49(15)(2021), 8974-8986.
21. Li S^#, Li X^#, Xue W^#, Zhang L^#, Yang LZ, Cao SM, Lei YN, Liu CX, Guo SK, Shan L, Wu M, Tao X, Zhang JL, Gao X, Zhang J, Wei J, Li J*, Yang L* & Chen L*. Screening for functional circular RNAs using the CRISPR–Cas13 system. Nat Methods 18(1)(2021), 51-59.
22. Wang Y*, Yan M*, Zhang X*, Liu X, Lai C, Ding Y, Tong MH* & Li J*. Rescue of male infertility through correcting a genetic mutation causing meiotic arrest in spermatogonial stem cells. Asian J Andrology 23(6)2021, 590-599.
23. Wang X^#, Kang JY^#, Wei L^#, Yang X^#, Sun H, Yang S, Liu L, Yan M, Bai M, Chen Y, Long J, Li N, Li D, Huang J, Lei M, Shao Z, Yuan W*, Zuo E*, Lu K*, Liu MF* & Li J*. PHF7 is a novel histone H2A E3 ligase prior to histone-to-protamine exchange during spermiogenesis. Development 146(13)(2019), dev175547.
24. Chen Y, Zheng Y, Gao Y, Lin Z, Yang S, Wang T, Wang Q, Xie N, Hua R, Liu M, Sha J, Griswold MD, Li J*, Tang F* & Tong M*. Single-cell RNA-seq uncovers dynamic processes and critical regulators in mouse spermatogenesis. Cell Res 28(9)(2018), 879-896.
25. Wang L^#, Li MY^#, Qu C, Miao WY, Yin Q, Liao J, Cao HT, Huang M, Wang K, Zuo E, Peng G, Zhang SX, Chen G, Li Q, Tang K, Yu Q, Li Z, Wong C, Xu G, Jing N, Yu X* & Li J*. CRISPR-Cas9-mediated genome-editing in one blastomere of two-cell embryos reveals a novel Tet3 function in regulating neocortical development. Cell Res 27(6)(2017), 815-829.
26. Wu K^#, Zhong C^#, Chen T^#, Zhang X, Tao W, Zhang J, Li H, Zhao H*, Li J* & Chen ZJ*. Polar bodies are efficient donors for reconstruction of human embryos for potential mitochondrial replacement therapy. Cell Res 27(8)(2017), 1069-1072.
27. Bai M, Liang D, Wang Y, Li Q, Wu Y & Li J*. Spermatogenic cell-specific gene mutation in mice via CRISPR-Cas9. J Genet Genomics 43(5)(2016), 289-296.
28. Wu Y^#, Zhou H^#, Fan X^#, Zhang Y^#, Zhang M^#, Wang Y, Xie Z, Bai M, Yin Q, Liang D, Tang W, Liao J, Zhou C, Liu W, Zhu P, Guo H, Pan H, Wu C, Shi H, Wu L*, Tang F* & Li J*. Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells. Cell Res 25(1)(2015), 67-79.
29. Zhang M, Zhou H, Zheng C, Xiao J, Zuo E, Liu W, Xie D, Shi Y, Wu C, Wang H, Li D & Li J*. The roles of testicular c-kit positive cells in de novo morphogenesis of testis. Sci Rep 4(2014), 5936. doi: 10.1038/srep05936.
30. Wu Y^#, Liang D^#, Wang Y, Bai M, Tang W, Bao S, Yan Z, Li D & Li J*. Correction of a genetic disease in mouse via use of CRISPR-Cas9. Cell Stem Cell 13(6)(2013), 659-662.
  - 三、细胞重编程相关论文（2007年—2013年）
31. Qin Y^#, Lin J^#, Zhou C^#, Yin Q, Xie Z, Zhang X, Liu X, Gao W & Li J*. Mice cloned from white adipose tissue-derived cells. J Mol Cell Biol 5(5)(2013), 348-350.
32. Guo F^#, Li X^#, Liang D^#, Li T^#, Zhu P, Guo H, Wu X, Wen L, Gu T, Hu B, Walsh CP, Li J*, Tang F* & Xu G*. Active and passive demethylation of male and female pronuclear DNA in the mammalian zygote. Cell Stem Cell 15(4)(2014), 447-458.
33. Jiang J^#, Lv W^#, Ye X, Wang L, Zhang M, Yang H, Okuka M, Zhou C, Zhang X, Liu L* & Li J*. Zscan4 promotes genomic stability during reprogramming and dramatically improves the quality of iPS cells as demonstrated by tetraploid complementation. Cell Res 23(1)(2013), 92-106.
34. Jiang J^#, Ding G^#, Lin J^#, Zhang M, Shi L, Lv W, Yang H, Xiao H, Pei G, Li Y, Wu J* & Li J*. Different developmental potential of pluripotent stem cells generated by different reprogramming strategies. J Mol Cell Biol 3(3)(2011), 197-199.
35. Gu T^#, Guo F^#, Yang H^#, Wu HP, Xu GF, Liu W, Xie ZG, Shi L, He X, Jin SG, Iqbal K, Shi YG, Deng Z, Szabo PE, Pfeifer GP, Li J* & Xu GL*. The role of Tet3 DNA dioxygenase in epigenetic reprogramming by oocytes. Nature 477(7366)(2011), 606-610.
36. Lin J, Shi L, Zhang M, Yang H, Qin Y, Zhang J, Gong D, Zhang X, Li D & Li J*. Defects in trophoblast cell lineage account for the impaired in vivo development of cloned embryos generated by somatic nuclear transfer. Cell Stem Cell 8(4)(2011), 371-375.
  - 四、应邀综述、评论（2012年—至今）
37. Li J* & Zeng Y*. Preface to the special topic on tissue stem cell research. Science China Life Science 62(6)(2021), 865-867. (Editoral)
38. Yan M & Li J*. Combined application of CRISPR-Cas9 and stem Cells for clinical and basic research. Cell Regeneration 9(2020), 19.
39. Yan M & Li J*. The evolving CRISPR technology. Protein Cell 10(11)(2019), 783-786.
40. Wang L & Li J*. Expansion of mutant monkey through cloning. Science China Life Science 62(6)(2019), 865-867. (Highlight)
41. Li Y & Li J*. Technical advances contribute to the study of genome imprinting. PLOS Genet 15(6)(2019), e1008151. (Invited review)
42. Wang L & Li J*. ‘Artificial spermatid’-mediated genome editing. Biol Reprod 101(3)(2019), 538-548. (Invited review)
43. Li J* & Gao C*. Preface to the special topic on genome editing research in China. Nat Sci Rev 6(3)(2019), 389-390.
44. Wang X & Li J*. An intermediate cell state allows rerouting of cell fate. J Biol Chem 292(46)(2017), 19133-19134. (Highlight).
45. Wang Y, Ding Y & Li J*. CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells. Methods Mol Biol 1622(2017), 293-305.
46. Zhong C & Li J*. Efficient generation of gene-modified mice by haploid embryonic stem cell-mediated semi-cloned technology. Methods Mol Biol 1498 (2017), 121-133.
47. Bai M, Wu Y & Li J*. Generation and application of mammalian haploid embryonic stem cells. J Intern Med 280(3)(2016), 236-245. (Invited Review)
48. Tong M* & Li J*. Similarity of epigenetic reprogramming in primordial germ cells between human and mouse. Natl Sci Rev 2(4)(2015), 384. (Invited Research Highlights)
49. Li J. Stem cell, basis and application. Sci Bull 60(20)(2015), 1711-1712.
50. Shi L, Yang H & Li J*. Haploid embryonic stem cells: an ideal tool for mammalian genetic analyses. Protein & Cell 3(11)(2012), 806-810. (Invited review)
  - 五、2007年以前的代表性论文
51. Li J & Mombaerts P. Nuclear transfer-mediated rescue of the nuclear genome of nonviable mouse cells frozen without cryoprotectant. Biol Reprod 79(4)(2008), 588-593.
52. Li J^#, Guasch G^#, Greco V^#, Fuchs E* & Mombaerts P*. Cloned mice from Skin Stem Cells. Proc Natl Acad Sci USA 104(8)(2007), 2738-2743.
53. Li J, Ishii T, Wen D & Mombaerts P. Non-equivalence of cloned and clonal mice. Curr Biol 15(18)(2005), R756-R757.
54. Li J, Ishii T, Feinstein P & Mombaerts P. Odorant receptor gene choice is reset by nuclear transfer from mouse olfactory sensory neurons. Nature 428(6981)(2004), 393-398.
55. Chen D^#, Li J^#, Han Z, Lei L, Liu Z, Kou Z, Ma S, Du Q & Sun Q. Somatic cell bovine cloning: efficiency of donor cells from female and male using different recipient cows. Chin Sci Bull 48(6)(2003), 549-554.

获奖及荣誉：

研究组成员：

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