CRISPR-Cas technologies have greatly reshaped the biology field.In this review,we discuss the CRISPR-Cas with a particular focus on the associated technologies and applications of CRISPR-Cas9 and CRISPR-Cas12a,which h...CRISPR-Cas technologies have greatly reshaped the biology field.In this review,we discuss the CRISPR-Cas with a particular focus on the associated technologies and applications of CRISPR-Cas9 and CRISPR-Cas12a,which have been most widely studied and used.We discuss the biological mechanisms of CRISPR-Cas as immune defense systems,recently-discovered anti-CRISPR-Cas systems,and the emerging Cas variants(such as xCas9 and Cas13)with unique characteristics.Then,we highlight various CRISPR-Cas biotechnologies,including nucleasedependent genome editing,CRISPR gene regulation(including CRISPR interference/activation),DNA/RNA base editing,and nucleic acid detection.Last,we summarize up-to-date applications of the biotechnologies for synthetic biology and metabolic engineering in various bacterial species.展开更多
Cas1 is a key component of the CRISPR adaptation complex,which captures and integrates foreign DNA into the CRISPR array,resulting in the generation of new spacers.We have determined crystal structures of Thermus ther...Cas1 is a key component of the CRISPR adaptation complex,which captures and integrates foreign DNA into the CRISPR array,resulting in the generation of new spacers.We have determined crystal structures of Thermus thermophilus Cas1 involved in new spacer acquisition both in complex with branched DNA and in the free state.Cas1 forms an asymmetric dimer without DNA.Conversely,two asymmetrical dimers bound to two branched DNAs result in the formation of a DNA-mediated tetramer,dimer of structurally asymmetrical dimers,in which the two subunits markedly present different conformations.In the DNA binding complex,the N-terminal domain adopts different orientations with respect to the C-terminal domain in the two monomers that form the dimer.Substrate binding triggers a conformational change in the loop 164–177 segment.This loop is also involved in the 3′fork arm and 5′fork arm strand recognition in monomer A and B,respectively.This study provides important insights into the molecular mechanism of new spacer adaptation.展开更多
Social dysfunction is a risk factor for several neuropsychiatric illnesses.Previous studies have shown that the lateral septum(LS)-related pathway plays a critical role in mediating social behaviors.Howeve r,the role ...Social dysfunction is a risk factor for several neuropsychiatric illnesses.Previous studies have shown that the lateral septum(LS)-related pathway plays a critical role in mediating social behaviors.Howeve r,the role of the connections between the LS and its downstream brain regions in social behavio rs remains unclea r.In this study,we conducted a three-chamber test using electrophysiological and chemogenetic approaches in mice to determine how LS projections to ventral CA1(vCA1)influence sociability.Our res ults showed that gamma-aminobutyric acid(GABA)-e rgic neuro ns were activated following social experience,and that social behavio rs were enhanced by chemogenetic modulation of these neurons.Moreover,LS GABAergic neurons extended their functional neural connections via vCA1 glutamatergic pyramidal neurons,and regulating LSGABA→vCA1Gluneural projections affected social behaviors,which were impeded by suppressing LSprojecting vCA1 neuronal activity or inhibiting GABAAreceptors in vCA1.These findings support the hypothesis that LS inputs to the vCA1 can control social prefe rences and social novelty behaviors.These findings provide new insights rega rding the neural circuits that regulate sociability.展开更多
蛋白磷酸酶是一组在人类细胞中广泛存在并催化已经磷酸化的蛋白质分子发生去磷酸化反应的一类酶分子。而磷酸蛋白磷酸酶催化亚基超家族是一组主要使丝氨酸/苏氨酸残基去磷酸化的蛋白磷酸酶,其中蛋白磷酸酶1(protein phosphatase-1,PP1)...蛋白磷酸酶是一组在人类细胞中广泛存在并催化已经磷酸化的蛋白质分子发生去磷酸化反应的一类酶分子。而磷酸蛋白磷酸酶催化亚基超家族是一组主要使丝氨酸/苏氨酸残基去磷酸化的蛋白磷酸酶,其中蛋白磷酸酶1(protein phosphatase-1,PP1)参与了真核生物大部分的去磷酸化反应,并参与调节多种生物活动。由PPP1CA基因编码的PP1催化亚基α(catalytic subunit alpha of protein phosphatase-1,PP1A)是修饰PP1最重要的一种亚型,在多种肿瘤生存和转移中起着重要作用,但其在肿瘤中也会表现出抑制作用,这使其可能成为肿瘤治疗的新靶标。因此,本文旨在对PP1A在多种肿瘤中关键作用进行综述,并针对其作为肿瘤治疗的新靶标潜力进行评价。展开更多
Dysregulation of hyperpolarization-activated cyclic nucleotide-gated cation(HCN)channels alters neuronal excitability.However,the role of HCN channels in status epilepticus is not fully understood.In this study,we est...Dysregulation of hyperpolarization-activated cyclic nucleotide-gated cation(HCN)channels alters neuronal excitability.However,the role of HCN channels in status epilepticus is not fully understood.In this study,we established rat models of pentylenetetrazole-induced status epilepticus.We performed western blot assays and immunofluorescence staining.Our results showed that HCN1 channel protein expression,particularly HCN1 surface protein,was significantly decreased in the hippocampal CA1 region,whereas the expression of HCN2 channel protein was unchanged.Moreover,metabolic glutamate receptor 1(mGluR1)protein expression was increased after status epilepticus.The mGluR1 agonist(RS)-3,5-dihydroxyphenylglycine injected intracerebroventricularly increased the sensitivity and severity of pentylenetetrazole-induced status epilepticus,whereas application of the mGluR1 antagonist(+)-2-methyl-4-carboxyphenylglycine(LY367385)alleviated the severity of pentylenetetrazole-induced status epilepticus.The results from double immunofluorescence labeling revealed that mGluR1 and HCN1 were co-localized in the CA1 region.Subsequently,a protein kinase A inhibitor(H89)administered intraperitoneally successfully reversed HCN1 channel inhibition,thereby suppressing the severity and prolonging the latency of pentylenetetrazole-induced status epilepticus.Furthermore,H89 reduced the level of mGluR1,downregulated cyclic adenosine monophosphate(cAMP)/protein kinase A expression,significantly increased tetratricopeptide repeat-containing Rab8b-interacting protein(TRIP8b)(1a-4)expression,and restored TRIP8b(1b-2)levels.TRIP8b(1a-4)and TRIP8b(1b-2)are subunits of Rab8b interacting protein that regulate HCN1 surface protein.展开更多
OBJECTIVE To investigate whether electroacupuncture(EA)ameliorates abnormal trigeminal neuralgia(TN)orofacial pain and anxiety-like behavior by altering synaptic plasticity in the hippocampus CA1.METHODS A mouse infra...OBJECTIVE To investigate whether electroacupuncture(EA)ameliorates abnormal trigeminal neuralgia(TN)orofacial pain and anxiety-like behavior by altering synaptic plasticity in the hippocampus CA1.METHODS A mouse infraorbital nerve transection model(pTION)of neuropathic pain was established,and EA or sham EA was used to treat ipsilateral acu⁃puncture points(GV20-Baihui and ST7-Xia⁃guan).Golgi-Cox staining and transmission elec⁃tron microscopy(TEM)were administrated to observe the changes of synaptic plasticity in the hippocampus CA1.RESULTS Stable and persistent orofacial allodynia and anxiety-like behav⁃iors induced by pT-ION were related to changes in hippocampal synaptic plasticity.Golgi stain⁃ings showed a decrease in the density of dendritic spines,especially mushroom-type dendritic spines,in hippocampal CA1 neurons of pT-ION mice.TEM results showed that the density of synapses,membrane thickness of the postsynaptic density,and length of the synaptic active zone were decreased,whereas the width of the synaptic cleft was increased in pTION mice.EA attenu⁃ated pT-ION-induced orofacial allodynia and anx⁃iety-like behaviors and effectively reversed the abnormal changes in dendritic spines and syn⁃apse of the hippocampal CA1 region.CONCLU⁃SION EA modulates synaptic plasticity of hippo⁃campal CA1 neurons,and reduces abnormal oro⁃facial pain and anxiety-like behavior,providing evidence for a TN treatment strategy.展开更多
基金Natural Science Foundation of Shanghai(Grant No.18ZR1420500)the Science and Technology Commission of Shanghai Municipality(Grant No.18JC1413600).Y.X.acknowledges National 1000 Youth Talents Program.W.L.and Y.Z.are recipients of China Postdoctoral Science Foundation(2018M632119 and 2018M632098).
文摘CRISPR-Cas technologies have greatly reshaped the biology field.In this review,we discuss the CRISPR-Cas with a particular focus on the associated technologies and applications of CRISPR-Cas9 and CRISPR-Cas12a,which have been most widely studied and used.We discuss the biological mechanisms of CRISPR-Cas as immune defense systems,recently-discovered anti-CRISPR-Cas systems,and the emerging Cas variants(such as xCas9 and Cas13)with unique characteristics.Then,we highlight various CRISPR-Cas biotechnologies,including nucleasedependent genome editing,CRISPR gene regulation(including CRISPR interference/activation),DNA/RNA base editing,and nucleic acid detection.Last,we summarize up-to-date applications of the biotechnologies for synthetic biology and metabolic engineering in various bacterial species.
基金The research was funded by the National Natural Science Foundation of China(31630015,31725008)the Chinese Academy of Sciences(QYZDY-SSW-SMC021).
文摘Cas1 is a key component of the CRISPR adaptation complex,which captures and integrates foreign DNA into the CRISPR array,resulting in the generation of new spacers.We have determined crystal structures of Thermus thermophilus Cas1 involved in new spacer acquisition both in complex with branched DNA and in the free state.Cas1 forms an asymmetric dimer without DNA.Conversely,two asymmetrical dimers bound to two branched DNAs result in the formation of a DNA-mediated tetramer,dimer of structurally asymmetrical dimers,in which the two subunits markedly present different conformations.In the DNA binding complex,the N-terminal domain adopts different orientations with respect to the C-terminal domain in the two monomers that form the dimer.Substrate binding triggers a conformational change in the loop 164–177 segment.This loop is also involved in the 3′fork arm and 5′fork arm strand recognition in monomer A and B,respectively.This study provides important insights into the molecular mechanism of new spacer adaptation.
基金supported by the National Natural Science Foundation of China,No.82171521(to CL)the Special Funds ofTaishan Scholars Project of Shandong Province,No.tsqn202211368(to CL)+2 种基金the Natural Science Foundation of Shandong Province,Nos.ZR2022YQ65(to CL),ZR2021MH073(to CL),ZR2019PH109(to WW)the Projects of Medical and Health Technology Development Program in Shandong Province,China,Nos.202003090720(to DZ),202003070728(to JL),2019 WS329(to DW)the Scientific Research Foundation of Binzhou Medical University,No.BY2018KJ21(to DW)。
文摘Social dysfunction is a risk factor for several neuropsychiatric illnesses.Previous studies have shown that the lateral septum(LS)-related pathway plays a critical role in mediating social behaviors.Howeve r,the role of the connections between the LS and its downstream brain regions in social behavio rs remains unclea r.In this study,we conducted a three-chamber test using electrophysiological and chemogenetic approaches in mice to determine how LS projections to ventral CA1(vCA1)influence sociability.Our res ults showed that gamma-aminobutyric acid(GABA)-e rgic neuro ns were activated following social experience,and that social behavio rs were enhanced by chemogenetic modulation of these neurons.Moreover,LS GABAergic neurons extended their functional neural connections via vCA1 glutamatergic pyramidal neurons,and regulating LSGABA→vCA1Gluneural projections affected social behaviors,which were impeded by suppressing LSprojecting vCA1 neuronal activity or inhibiting GABAAreceptors in vCA1.These findings support the hypothesis that LS inputs to the vCA1 can control social prefe rences and social novelty behaviors.These findings provide new insights rega rding the neural circuits that regulate sociability.
文摘蛋白磷酸酶是一组在人类细胞中广泛存在并催化已经磷酸化的蛋白质分子发生去磷酸化反应的一类酶分子。而磷酸蛋白磷酸酶催化亚基超家族是一组主要使丝氨酸/苏氨酸残基去磷酸化的蛋白磷酸酶,其中蛋白磷酸酶1(protein phosphatase-1,PP1)参与了真核生物大部分的去磷酸化反应,并参与调节多种生物活动。由PPP1CA基因编码的PP1催化亚基α(catalytic subunit alpha of protein phosphatase-1,PP1A)是修饰PP1最重要的一种亚型,在多种肿瘤生存和转移中起着重要作用,但其在肿瘤中也会表现出抑制作用,这使其可能成为肿瘤治疗的新靶标。因此,本文旨在对PP1A在多种肿瘤中关键作用进行综述,并针对其作为肿瘤治疗的新靶标潜力进行评价。
基金supported by the National Natural Science Foundation of China,No.81760242(to MGM).
文摘Dysregulation of hyperpolarization-activated cyclic nucleotide-gated cation(HCN)channels alters neuronal excitability.However,the role of HCN channels in status epilepticus is not fully understood.In this study,we established rat models of pentylenetetrazole-induced status epilepticus.We performed western blot assays and immunofluorescence staining.Our results showed that HCN1 channel protein expression,particularly HCN1 surface protein,was significantly decreased in the hippocampal CA1 region,whereas the expression of HCN2 channel protein was unchanged.Moreover,metabolic glutamate receptor 1(mGluR1)protein expression was increased after status epilepticus.The mGluR1 agonist(RS)-3,5-dihydroxyphenylglycine injected intracerebroventricularly increased the sensitivity and severity of pentylenetetrazole-induced status epilepticus,whereas application of the mGluR1 antagonist(+)-2-methyl-4-carboxyphenylglycine(LY367385)alleviated the severity of pentylenetetrazole-induced status epilepticus.The results from double immunofluorescence labeling revealed that mGluR1 and HCN1 were co-localized in the CA1 region.Subsequently,a protein kinase A inhibitor(H89)administered intraperitoneally successfully reversed HCN1 channel inhibition,thereby suppressing the severity and prolonging the latency of pentylenetetrazole-induced status epilepticus.Furthermore,H89 reduced the level of mGluR1,downregulated cyclic adenosine monophosphate(cAMP)/protein kinase A expression,significantly increased tetratricopeptide repeat-containing Rab8b-interacting protein(TRIP8b)(1a-4)expression,and restored TRIP8b(1b-2)levels.TRIP8b(1a-4)and TRIP8b(1b-2)are subunits of Rab8b interacting protein that regulate HCN1 surface protein.
基金the National Natural Science Foundation of China(82001190)Natural Sci⁃ence Foundation of Shandong Province(ZR2021LZY016)+1 种基金Natural Science Foundation of Shandong Province(ZR2020MH348)Science and Technology Foundation of Shandong Traditional Chinese Medicine(2020Q035)。
文摘OBJECTIVE To investigate whether electroacupuncture(EA)ameliorates abnormal trigeminal neuralgia(TN)orofacial pain and anxiety-like behavior by altering synaptic plasticity in the hippocampus CA1.METHODS A mouse infraorbital nerve transection model(pTION)of neuropathic pain was established,and EA or sham EA was used to treat ipsilateral acu⁃puncture points(GV20-Baihui and ST7-Xia⁃guan).Golgi-Cox staining and transmission elec⁃tron microscopy(TEM)were administrated to observe the changes of synaptic plasticity in the hippocampus CA1.RESULTS Stable and persistent orofacial allodynia and anxiety-like behav⁃iors induced by pT-ION were related to changes in hippocampal synaptic plasticity.Golgi stain⁃ings showed a decrease in the density of dendritic spines,especially mushroom-type dendritic spines,in hippocampal CA1 neurons of pT-ION mice.TEM results showed that the density of synapses,membrane thickness of the postsynaptic density,and length of the synaptic active zone were decreased,whereas the width of the synaptic cleft was increased in pTION mice.EA attenu⁃ated pT-ION-induced orofacial allodynia and anx⁃iety-like behaviors and effectively reversed the abnormal changes in dendritic spines and syn⁃apse of the hippocampal CA1 region.CONCLU⁃SION EA modulates synaptic plasticity of hippo⁃campal CA1 neurons,and reduces abnormal oro⁃facial pain and anxiety-like behavior,providing evidence for a TN treatment strategy.
