干细胞耗竭是引起机体衰老的重要原因，细胞衰老过程中核膜异常和核纤层蛋白表达下调，说明核膜蛋白在调控细胞衰老方面扮演重要的作用，但核膜蛋白如何调节人干细胞稳态和衰老尚不清楚，我们在人胚胎干细胞中利用CRISPR/Cas9技术将核膜蛋白基因LMNA、LMNB1、LMNB2、EMD进行敲除，并将获得的敲除细胞系定向分化成人不同类型的成体干细胞和终末分化细胞，观察核膜蛋白缺失对细胞增殖和衰老的影响，对异染色质稳定性的影响，观察核膜蛋白对衰老的调控是否存在冗余性，通过RNA-seq、ChIP-seq、DamID-seq和ATAT-seq综合分析，探究核膜蛋白调节衰老的分子机制，通过药物筛选寻找干预靶点，为延缓干细胞衰老提供线索。

Stem cell depletion is an important element of aging．In the aging cell, nuclear membrane become irregular and lamin is down-regulated, which reveal nuclear envelope proteins play an important role during aging. In order to research the important function of the nuclear envelope proteins in maintaining stem cells homeostasis, and to study how nuclear envelope proteins mediate the stability of the heterochromatin, we generated LMNA, LMNB1,LMNB2,EMD-deficient human embryonic stem cells (hESCs) by using CRISPR/Cas9 technique. After inducing differentiation, nuclear envelope proteins deficient human mesenchymal stem cells (hMSCs) exhibited delayed proliferation rate and premature senescence. On other hand, regulation mechanism of Nuclear membrane proteins to the cell aging will be studied by RNA-seq, ChIP-seq and ATAT-seq. In addition, drug screening will be used to find the target of intervention and provide clues for delaying the senescence of stem cells.