Subculturing cells have no effect on CRISPR/Cas9-mediated cleavage of UL30 gene in pseudorabies virus

CRISPR/Cas9-mediated genome editing can inhibit virus infection by targeting the conserved regions of the viral genomic DNA. Unexpectedly, we found previously that pseudorabies virus(PRV) could escape from CRISPR/Cas9-mediated inhibition.In order to elucidate whether the escape of PRV from Cas9-mediated inhibition was due to cell deficiencies, such as genetic instability of sgRNA or Cas9 protein, the positive cells were passaged ten times, and PRV infection in the sgRNA-expressing cells was evaluated in the present study. The results showed that subculturing cells has no effect on Cas9-mediated cleavage of PRV. Different passages of PX459-PRV cells can stably express sgRNA to facilitate Cas9/sgRNA cleavage on the UL30 gene of PRV, resulting in a pronounced inhibition of PRV infection. Studies to elucidate the mechanism of PRV escape are currently in progress.

Studies of an X-ray imaging detector based on THGEM and CCD camera

Introduction The photons generated by the electron avalanche in gaseous detectors are known as the secondary scintillation light,of which the spectrum range is from ultraviolet to visible.So it is possible to collect directly the avalanche-induced photons in visible range by light sensors,such as charged-couple device(CCD).The optical readout is a new method for 2-D imaging of high spatial resolution based on the micro-pattern gaseous detectors(MPGDs).Purpose The traditional charge readout method for the MPGDs is complicated,expensive,and strongly depends on the technology of application-specific integrated circuit.In some cases,for example,low-frame-rate 2-D imaging,the optical readout has more advantages,such as simple,cheap and easy to use.Therefore,a gaseous detector was developed based on the thick gaseous electron multiplier(THGEM)and the ordinary scientific CCD camera as readout to realize the X-ray imaging instead of charge readout.Methods and materials The THGEMs with 60×60 mm^(2) sensitive area were developed to reach high gain,and the Ar+CF4 gas mixture was chosen for strong light emission.And so a general scientific CCD can be used for readout and replacing the intensified CCD,which is more expensive than general CCD.Results Some clear X-ray images were obtained by this optical readout THGEM detector,and the spatial resolution achieved is 275µm.It is indicated that this kind of detectors have promising imaging capability and great potential for practical application.

A new neutron detector based on ceramic THGEM and boron-coated meshes

Introduction Boron-coated micro-pattern gaseous detectors,such as gaseous electron multiplier(GEM),are widely studied as the neutron detectors.The existing solution to improve the neutron detection efficiency(NDE)is to coat the boron on the surfaces of the cascade GEM films.So the GEM films themselves will be affected by the complicated coating process and the boron layers,and the 10B material will be wasted once the GEM films are damaged.Since the neutron conversion and the electron multiplication regions are not separated,the final gains of different layer ionized electrons are very difficult to control and keep consistent.Materials and methods A new neutron detector based on the ceramic thick GEM(THGEM)was proposed.The boron-coated cathode and multilayer meshes were used as the neutron conversion region and separated from the electron multiplication region consisting of only one THGEM film.The sawtooth surface treatment and the double-sided readout method were applied to improve neutron conversion efficiency(NCE).The neutron conversion and the electron collection efficiencies were simulated and studied based on the new structure.The prototype detector was test by using a ^(252)Cf neutron source.Results The simulation results show that the cascade boron-coated meshes have good neutron conversion and electron collection efficiencies,and the double-sided readout and sawtooth structure can greatly increase the NCE.The neutron test results show that the detection efficiency of the double-sided readout is 1.6 times higher than that of the common single-sided readout.These indicate that the new detector has high neutron conversion and detection efficiencies and is promising for practical applications.

A scintillator detector for beam tuning of low-energy single electron accelerator

Purpose Nine scintillator detectors were developed for beam tuning of low-energy single electron accelerator instead of the picoammeter and Faraday cup,both of which will be insensitive to the weak-current beam lower than picoampere.Methods The detectors are based on plastic scintillator and photomultiplier tube(PMT)and the beam is 100 Hz pulsed electrons in the energy range of 0.1 to 50MeV.The beam flux was tuned from about 107 to quasi-single electron for each bunch according to the measurement results of these detectors installed along the beam pipe and after slits.In order to determine the number of electrons in a bunch,the gains of each PMT were finely calibrated,and the energy response of each detector was calibrated with 207Bi radioactive source.Moreover,the deposited energy in the plastic scintillator,the electron detection efficiency and light collection efficiency,as a function of electron incident energy,were also simulated in detail by Geant4.Results Nine detectors worked well at a gain of 0.6×10^(6) after installation in the beam pipe,and the number of electrons in a bunch could be measured at each detector position.The single electron peak was seen when the beam reached the final detector.Conclusion The commissioning result shows that the beam was tuned to quasi-single electron level successfully and the scintillator detector is an effective method to measure the weak-current beam lower than picoampere.