Construction and characterization of a synthesized herpes simplex virus H129-Syn-G2

Herpes simplex virus type 1(HSV-1)causes lifelong infections worldwide,and currently there is no efficient cure or vaccine.HSV-1-derived tools,such as neuronal circuit tracers and oncolytic viruses,have been used exten-sively;however,further genetic engineering of HSV-1 is hindered by its complex genome structure.In the present study,we designed and constructed a synthetic platform for HSV-1 based on H129-G4.The complete genome was constructed from 10 fragments through 3 rounds of synthesis using transformation-associated recombination(TAR)in yeast,and was named H129-Syn-G2.The H129-Syn-G2 genome contained two copies of the gfp gene and was transfected into cells to rescue the virus.According to growth curve assay and electron microscopy results,the synthetic viruses exhibited more optimized growth properties and similar morphogenesis compared to the parental virus.This synthetic platform will facilitate further manipulation of the HSV-1 genome for the devel-opment of neuronal circuit tracers,oncolytic viruses,and vaccines.

Multigene phylogeny of the family Cordycipitaceae (Hypocreales): new taxa and the new systematic position of the Chinese cordycipitoid fungus Paecilomyces hepiali

The phylogeny and systematics of cordycipitoid fungi have been extensively studied in the last two decades.However,systematic positions of some taxa in the family Cordycipitaceae have not yet been thoroughly resolved.In this study,a new phylogenetic framework of Cordycipitaceae is reconstructed using multigene(nrSSU,nrLSU,tef-1α,rpb1 and rpb2)sequence data with large-scale taxon sampling.In addition,ITS sequence data of species belonging to the Lecanicillium lineage in the family Cordycipitaceae are used to further determine their phylogenetic placements.Based on molecular phylogenetic data together with morphological evidence,two new genera(Flavocillium and Liangia),16 new species and four new combina-tions are introduced.In the new genus Flavocillium,one new species F.bifurcatum and three new combinations previously described as Lecanicillium,namely F.acerosium,F.primulinium and F.subprimulinium,are proposed.The genus Liangia is built by the new species Lia.sinensis with Lecanicillium-like asexual morph,isolated from an entomopathogenic fungus Beauveria yunnanensis.Due to the absence of Paecilomyces hepiali,an economically and medically significant fungus,in the earlier phylogenetic analyses,its systematic position has been puzzling in both business and academic communities for a long time.Here,P.hepiali is recharacterized using the holotype material along with seven additional samples.It is assigned to the genus Samsoniella(Cordycipitaceae,Hypocreales)possessing Cordyceps-like sexual morph and Isaria-like asexual morph,and thus a new combination,namely S.hepiali is proposed.An additional nine new species in Samsoniella are described:S.alpina,S.antleroides,S.cardinalis,S.cristata,S.lanmaoa,S.kunmingensis,S.ramosa,S.tortricidae and S.yunnanensis.Four new species in Cordyceps are described:C.chaetoclavata,C.cocoonihabita,C.shuifuensis and C.sub-tenuipes.Simplicillium yunnanense,isolated from synnemata of Akanthomyces waltergamsii,is described as a new species.

Synthesis and characterization of star-branched polyamide 6 via anionic ring-opening polymerization with N,N',N''-trimesoyltricaprolactam as a multifunctional activator

Star-branched polyamide 6 was prepared via anionic ring-opening polymerization of e-caprolactam in the presence of a simple benzene-centered trifunctional activator of N,N',N''-trimesoyltricaprolactam. A high polymer yields of above 95% were achieved at 160 8C for 15 min utilizing e-caprolactam magnesium bromide as a catalyst. Compared with its linear counterpart, the resultant star-branched polyamide 6showed smaller relative viscosity(1.51), decreased melting temperature(218 8C) and lower crystallinity(24.2%). The specific properties demonstrated the existence of a star-branched structure and provided potential advantages in engineering applications.

Anterograde Trans-Synaptic Tagging Mediated by Adeno-Associated Virus

Trans-neuronal viral tracing is becoming one of the most important methods for mapping neuronal circuit connections,but an anterograde trans-synaptic tracer compatible with functional assays is still lacking.Adeno-associated virus(AAV)is replication-defective,and its production of offspring needs the help of adenovirus(AV)[1]or herpesvirus co-infection[2].The replication deficiency,non-pathogenicity,easy manipulability。

HSV-1 H129-Derived Anterograde Neural Circuit Tracers:Improvements, Production, and Applications

Anterograde viral tracers are powerful and essential tools for dissecting the output targets of a brain region of interest. They have been developed from herpes simplex virus 1(HSV-1) strain H129(H129), and have been successfully applied to map diverse neural circuits.Initially, the anterograde polysynaptic tracer H129-G4 was used by many groups. We then developed the first monosynaptic tracer, H129-dTK-tdT, which was highly successful, yet improvements are needed. Now, by inserting another tdTomato expression cassette into the H129-dTK-tdT genome, we have created H129-dTK-T2, an updated version of H129-dTK-tdT that has improved labeling intensity. To help scientists produce and apply our H129-derived viral tracers, here we provide the protocol describing our detailed and standardized procedures. Commonly-encountered technical problems and their solutions are also discussed in detail. Broadly, the dissemination of this protocol will greatly support scientists to apply these viral tracers on a large scale.

Concerns on Vaccine against Varicella Caused by Varicella-Zoster Virus Infection

Varicella-zoster virus(VZV)belongs to a neurotropic Alphaherpesvirinae and is the causative pathogen of both the varicella and herpes zoster.Currently,there are three different vaccines for the prevention,low does live-attenuated Oka(v Oka)against varicella,high does v Oka and glycoprotein E with the AS01B adjuvant system against zoster.Varicella vaccine containing v Oka effectively prevents varicella,but latency of v Oka can be detected in the inoculated population and its reactivation causes zoster.

Human cytomegalovirus infection dysregulates neural progenitor cell fate by disrupting Hes1 rhythm and down-regulating its expression

Human cytomegalovirus(HCMV) infection is a leading cause of birth defects, primarily affecting the central nervous system and causing its maldevelopment. As the essential downstream effector of Notch signaling pathway, Hes1, and its dynamic expression, plays an essential role on maintaining neural progenitor/stem cells(NPCs) cell fate and fetal brain development. In the present study, we reported the first observation of Hes1 oscillatory expression in human NPCs, with an approximately1.5 hour periodicity and a Hes1 protein half-life of about 17(17.6 ± 0.2) minutes. HCMV infection disrupts the Hes1 rhythm and down-regulates its expression. Furthermore, we discovered that depleting Hes1 protein disturbed NPCs cell fate by suppressing NPCs proliferation and neurosphere formation, and driving NPCs abnormal differentiation. These results suggested a novel mechanism linking disruption of Hes1 rhythm and down-regulation of Hes1 expression to neurodevelopmental disorders caused by congenital HCMV infection.