CMB Research Support Specialists provide technical expertise and support to NIAID Principal Investigators, assisting them with true cage to benchside support. Both pathology and technical proficiency resulted in numerous co-authorship opportunities. The CMB has successfully maintained a gnotobiotic breeding and study facility, which consists of bio-exclusion units designed to keep the mice from becoming colonized with any adventitious microorganisms. Germ-free mice are free of all aerobic and anaerobic organisms with the possible exception of endogenous viruses. Breeding colonies and mice on study are maintained in isolators provided with HEPA-filtered air and autoclaved food, bedding, and supplies. Strict SOPs are followed to maintain the mice in a germ-free state. The Mouse Genetics and Gene Modification (MGGM) Section at CMB/NIAID provides highly sophisticated CRISPR Cas9 mediated genome editing in embryos and in cells as well as related services such as; a) cryopreservation of sperm; b) cryopreservation of embryo for long term storage of valuable mouse lines and c) rederivation of lines from cryopreserved embryos and sperm. New systems such as, ss-oligonucleotides (ss-ODNs) and large DNA plasmids to create (KI) and cre-lox knockout (KO) mice were tested. More than 45 NIAID investigators used MGGM services for the CRISPR gene KO/KI, sperm cryopreservation, embryo cryopreservation and rederivation of lines. CRISPR cas9 mediated genome editing: Gene KO by CRISPR/cas9 was completed for 6 projects and 5 projects were completed for gene KI. In addition, 30-50 pups were generated for 6 gene KI projects involving large plasmids and cre-lox for conditional gene KO. 2 CRISPR/cas9 gene projects were also completed KO in BV2 and THP1 somatic cells. Three publications have resulted from the efforts from MGGM work in collaboration with NIAID and other NIH investigators. Cryopreservation of mouse lines via cryopreservation of sperm and embryo and rederivation of lines : a) Successfully completed 98 sperm cryopreservation projects; b) 49 of which were tested for sperm QC; c) 13 embryo cryo projects were completed; d) 8 embryo cryo projects were tested for QC by creating live pups; e) A total of 31 projects were completed for rederivation of lines with the embryos either cryopreserved by MGGM or imported from outside sources. The Infectious Disease Pathogenesis Section (IDPS) supports NIAID investigators in the execution of many activities related to animal studies including, but not limited to: animal study design; necropsy instruction and assistance; creating and refining tissue sample collection protocols; ensuring that investigators are well-informed of animal model-related anatomy, physiology, and appropriate pathology terminology. The IDPS also aids in the interpretation of data and the incorporation of these data into manuscripts for publication. The IDPS continues to improve the quality of pathology support given to NIAID investigators and the level of communication/collaboration between the IDPS and the laboratories within NIAID. The IDPS has made a number of functional changes and improvements to the services provided to the NIAID investigator community. The IDPS has recently transitioned to a full-service histopathology laboratory with full tissues-in-slides-out capabilities. In addition, the IDPS has acquired a state-of-the-art digital pathology slide scanning system that allows easily sharing of data both internally and abroad and facilitates performing semi-quantitative image analysis on a diverse set of samples. These technologies will now give the IDPS the ability to process a wide range of tissue types and perform various analyses within our laboratory space. To date, the IDPS continues to generate traditional and innovative pathology data sets for investigators; many of these data sets are associated with publications that are currently in progress, have been submitted for publication or, have been accepted in highly respected, peer-reviewed, journals including Nature, Nature Immunology, Plos Pathogens, and Journal of Infectious Diseases. Our main objectives are to continually increase the laboratorys efficiency and scientific contribution without compromising quality; retain the IDPS quest for innovation by continued implementation of cutting-edge technology; and uphold a commitment to maintaining the highest standard of pathology research support for the NIAID investigator community. The IDPS most significant contributions have, in large, been related to our ability to help highlight the pertinent scientific details that reside in the gap between the generation of a hypothesis and the benchtop. We help investigators examine the broader and more basic pathogenesis of less well-characterized disease processes and also help clarify the significance of newly identified disease-specific features in animal model systems.
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