Intracranial aneurysm(IA)is characterized by defects in the middle muscular layer and pathological dilatation of cerebral arteries.The rupture of IA,resulting in aneurysmal subarachnoid hemorrhage,poses a substantial ...Intracranial aneurysm(IA)is characterized by defects in the middle muscular layer and pathological dilatation of cerebral arteries.The rupture of IA,resulting in aneurysmal subarachnoid hemorrhage,poses a substantial public health problem,especially in developed countries,due to its high mortality,morbidity,and socioeconomic burden(Bakker and Ruigrok,2021).Understanding the risk factors for IA is critical for disease prevention.Although numerous studies(Yan et al.,2015;Santiago-Sim et al.,2016;Barak et al.,2021;Liu et al.,2021)have highlighted familial aggregation of IA and identified candidate genes such as ADAMTS15,THSD1,ANK3,and PPIL4,the mechanisms underlying IA formation and rupture remain unclear.展开更多
Mycobacterium tuberculosis is the causative agent of tuberculosis(TB), which is still the leading cause of mortality from a single infectious disease worldwide. The development of novel anti-TB drugs and vaccines is s...Mycobacterium tuberculosis is the causative agent of tuberculosis(TB), which is still the leading cause of mortality from a single infectious disease worldwide. The development of novel anti-TB drugs and vaccines is severely hampered by the complicated and time-consuming genetic manipulation techniques for M. tuberculosis. Here, we harnessed an endogenous type Ⅲ-A CRISPR/Cas10 system of M. tuberculosis for efficient gene editing and RNA interference(RNAi).This simple and easy method only needs to transform a single mini-CRISPR array plasmid, thus avoiding the introduction of exogenous protein and minimizing proteotoxicity. We demonstrated that M. tuberculosis genes can be efficiently and specifically knocked in/out by this system as confirmed by DNA high-throughput sequencing. This system was further applied to single-and multiple-gene RNAi. Moreover, we successfully performed genome-wide RNAi screening to identify M. tuberculosis genes regulating in vitro and intracellular growth. This system can be extensively used for exploring the functional genomics of M. tuberculosis and facilitate the development of novel anti-TB drugs and vaccines.展开更多
The human genome contains around 20,000 protein-coding genes and a vast of transcriptional regulatory elements(ENCODE Project Consortium,2012;Roadmap Epigenomics Consortium et al.,2015;Stunnenberg and Hirst,2016).It s...The human genome contains around 20,000 protein-coding genes and a vast of transcriptional regulatory elements(ENCODE Project Consortium,2012;Roadmap Epigenomics Consortium et al.,2015;Stunnenberg and Hirst,2016).It still remains elusive how these regulatory elements regulate their target genes across genomic distances of millions of bases.展开更多
基金supported by the National Natural Science Foundation of China(32222028)Natural Science Foundation of Chongqing,China(CSTB2023NSCQ-MSX0505)+1 种基金Financial Science and Technology Project of Hunan Province,China(422000008)Hunan Province Nature Science Foundation,China(2021JJ30911).
文摘Intracranial aneurysm(IA)is characterized by defects in the middle muscular layer and pathological dilatation of cerebral arteries.The rupture of IA,resulting in aneurysmal subarachnoid hemorrhage,poses a substantial public health problem,especially in developed countries,due to its high mortality,morbidity,and socioeconomic burden(Bakker and Ruigrok,2021).Understanding the risk factors for IA is critical for disease prevention.Although numerous studies(Yan et al.,2015;Santiago-Sim et al.,2016;Barak et al.,2021;Liu et al.,2021)have highlighted familial aggregation of IA and identified candidate genes such as ADAMTS15,THSD1,ANK3,and PPIL4,the mechanisms underlying IA formation and rupture remain unclear.
基金supported by the National Key R&D Program of China(Grant No.2017YFD0500303)the National Natural Science Foundation of China(Grant Nos.C180501 and 31602061)+1 种基金the Huazhong Agricultural University Scientific&Technological Self-innovation Foundation,China(Grant Nos.2662017PY105 and 2662017PY105)the Doctoral Fund of Ministry of Education of China(Grant No.131012).
文摘Mycobacterium tuberculosis is the causative agent of tuberculosis(TB), which is still the leading cause of mortality from a single infectious disease worldwide. The development of novel anti-TB drugs and vaccines is severely hampered by the complicated and time-consuming genetic manipulation techniques for M. tuberculosis. Here, we harnessed an endogenous type Ⅲ-A CRISPR/Cas10 system of M. tuberculosis for efficient gene editing and RNA interference(RNAi).This simple and easy method only needs to transform a single mini-CRISPR array plasmid, thus avoiding the introduction of exogenous protein and minimizing proteotoxicity. We demonstrated that M. tuberculosis genes can be efficiently and specifically knocked in/out by this system as confirmed by DNA high-throughput sequencing. This system was further applied to single-and multiple-gene RNAi. Moreover, we successfully performed genome-wide RNAi screening to identify M. tuberculosis genes regulating in vitro and intracellular growth. This system can be extensively used for exploring the functional genomics of M. tuberculosis and facilitate the development of novel anti-TB drugs and vaccines.
基金supported by the National Natural Science Foundation of China(31900432 to D.L.,31771402 to G.L.)the National Postdoctoral Program for Innovative Talents(BX20180113 to D.L.)+1 种基金China Postdoctoral Science Foundation(2018M640710 to D.L.)the Fundamental Research Funds for the Central Universities(2662018PY025 to G.C.)
文摘The human genome contains around 20,000 protein-coding genes and a vast of transcriptional regulatory elements(ENCODE Project Consortium,2012;Roadmap Epigenomics Consortium et al.,2015;Stunnenberg and Hirst,2016).It still remains elusive how these regulatory elements regulate their target genes across genomic distances of millions of bases.
