@article{klotz_goh_o'flaherty_johnson_barrangou_2020, title={Deletion of S-Layer Associated Ig-Like Domain Protein Disrupts the Lactobacillus acidophilus Cell Surface}, volume={11}, ISSN={["1664-302X"]}, DOI={10.3389/fmicb.2020.00345}, abstractNote={Bacterial surface-layers (S-layers) are crystalline arrays of repeating proteinaceous subunits that coat the exterior of many cell envelopes. S-layers have demonstrated diverse functions in growth and survival, maintenance of cell integrity, and mediation of host interactions. Additionally, S-layers can act as scaffolds for the outward display of auxiliary proteins and glycoproteins. These non-covalently bound S-layer associated proteins (SLAPs) have characterized roles in cell division, adherence to intestinal cells, and modulation of the host immune response. Recently, IgdA (LBA0695), a Lactobacillus acidophilus SLAP that possesses a Group 3 immunoglobulin (Ig)-like domain and GW (Gly-Tryp) dipeptide surface anchor, was recognized for its high conservation among S-layer-forming lactobacilli, constitutive expression, and surface localization. These findings prompted its selection for examination within the present study. Although IgdA and corresponding orthologs were shown to be unique to host-adapted lactobacilli, the Ig domain itself was specific to vertebrate-adapted species suggesting a role in vertebrate adaptation. Using a counterselective gene replacement system, igdA was deleted from the L. acidophilus NCFM chromosome. The resultant mutant, NCK2532, exhibited a visibly disrupted cell surface which likely contributed to its higher salt sensitivity, severely reduced adhesive capacity, and altered immunogenicity profile. Transcriptomic analyses revealed the induction of several stress response genes and secondary surface proteins. Due to the broad impact of IgdA on the cellular physiology and probiotic attributes of L. acidophilus, identification of similar proteins in alternative bacterial species may help pinpoint next-generation host-adapted probiotic candidates.}, journal={FRONTIERS IN MICROBIOLOGY}, author={Klotz, Courtney and Goh, Yong Jun and O'Flaherty, Sarah and Johnson, Brant and Barrangou, Rodolphe}, year={2020}, month={Mar} }
 @article{klotz_2019, title={CRISPR Cinema: Scenes from the Cutting Edge}, volume={2}, ISSN={["2573-1602"]}, DOI={10.1089/crispr.2019.29056.ckl}, abstractNote={The CRISPR JournalVol. 2, No. 2 First CutsCRISPR Cinema: Scenes from the Cutting EdgeCourtney KlotzCourtney KlotzAddress correspondence to: Courtney Klotz,North Carolina State University, Raleigh, NC,E-mail Address: ceklotz@ncsu.eduNorth Carolina State University, Raleigh, North CarolinaSearch for more papers by this authorPublished Online:18 Apr 2019https://doi.org/10.1089/crispr.2019.29056.cklAboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View articleFiguresReferencesRelatedDetailsCited byDrug discovery: Chaos can be your friend or your enemyThe Director's Cut: An Interview with Adam Bolt Kevin Davies and Adam Bolt16 August 2019 | The CRISPR Journal, Vol. 2, No. 4CRISPR-Accelerated Gene Drives Pump the Brakes Julianna LeMieux16 August 2019 | The CRISPR Journal, Vol. 2, No. 4 Volume 2Issue 2Apr 2019 InformationCopyright 2019, Mary Ann Liebert, Inc.To cite this article:Courtney Klotz.CRISPR Cinema: Scenes from the Cutting Edge.The CRISPR Journal.Apr 2019.76-78.http://doi.org/10.1089/crispr.2019.29056.cklPublished in Volume: 2 Issue 2: April 18, 2019PDF download}, number={2}, journal={CRISPR JOURNAL}, author={Klotz, Courtney}, year={2019}, month={Apr}, pages={76–78} }
 @misc{klotz_barrangou_2018, title={Engineering Components of the Lactobacillus S-Layer for Biotherapeutic Applications}, volume={9}, ISSN={["1664-302X"]}, DOI={10.3389/fmicb.2018.02264}, abstractNote={Lactic acid bacteria (LAB) are frequently harnessed for the delivery of biomolecules to mucosal tissues. Several species of Lactobacillus are commonly employed for this task, of which a subset are known to possess surface-layers (S-layers). S-layers are two-dimensional crystalline arrays of repeating proteinaceous subunits that form the outermost coating of many prokaryotic cell envelopes. Their periodicity and abundance have made them a target for numerous biotechnological applications. In the following review, we examine the multi-faceted S-layer protein (Slp), and its use in both heterologous protein expression systems and mucosal vaccine delivery frameworks, through its diverse genetic components: the strong native promoter, capable of synthesizing as many as 500 Slp subunits per second; the signal peptide that stimulates robust secretion of recombinant proteins; and the structural domains, which can be harnessed for both cell surface display of foreign peptides or adhesion enhancement of a host bacterium. Although numerous studies have established vaccine platforms based on one or more components of the Lactobacillus S-layer, this area of research still remains largely in its infancy, thus this review is meant to not only highlight past works, but also advocate for the future usage of Slps in biotherapeutic research.}, journal={FRONTIERS IN MICROBIOLOGY}, publisher={Frontiers Media SA}, author={Klotz, Courtney and Barrangou, Rodolphe}, year={2018}, month={Oct} }
 @article{griffin_witrick_klotz_dorenkott_goodrich_fundaro_mcmillan_hulver_ponder_neilson_2017, title={Alterations to metabolically active bacteria in the mucosa of the small intestine predict anti-obesity and anti-diabetic activities of grape seed extract in mice}, volume={8}, ISSN={["2042-650X"]}, DOI={10.1039/c7fo01236e}, abstractNote={Grape seed extract changes small intestinal gut microbiota composition.}, number={10}, journal={FOOD & FUNCTION}, author={Griffin, Laura E. and Witrick, Katherine A. and Klotz, Courtney and Dorenkott, Melanie R. and Goodrich, Katheryn M. and Fundaro, Gabrielle and McMillan, Ryan P. and Hulver, Matthew W. and Ponder, Monica A. and Neilson, Andrew P.}, year={2017}, month={Oct}, pages={3510–3522} }
 @article{klotz_o'flaherty_goh_barrangou_2017, title={Investigating the Effect of Growth Phase on the Surface-Layer Associated Proteome of Lactobacillus acidophilus Using Quantitative Proteomics}, volume={8}, ISSN={["1664-302X"]}, DOI={10.3389/fmicb.2017.02174}, abstractNote={Bacterial surface-layers (S-layers) are semi-porous crystalline arrays that self-assemble to form the outermost layer of some cell envelopes. S-layers have been shown to act as scaffolding structures for the display of auxiliary proteins externally. These S-layer associated proteins have recently gained attention in probiotics due to their direct physical contact with the intestinal mucosa and potential role in cell proliferation, adhesion, and immunomodulation. A number of studies have attempted to catalog the S-layer associated proteome of Lactobacillus acidophilus NCFM under a single condition. However, due to the versatility of the cell surface, we chose to employ a multiplexing-based approach with the intention of accurately contrasting multiple conditions. In this study, a previously described lithium chloride isolation protocol was used to release proteins bound to the L. acidophilus S-layer during logarithmic and early stationary growth phases. Protein quantification values were obtained via TMT (tandem mass tag) labeling combined with a triple-stage mass spectrometry (MS3) method. Results showed significant growth stage-dependent alterations to the surface-associated proteome while simultaneously highlighting the sensitivity and reproducibility of the technology. Thus, this study establishes a framework for quantifying condition-dependent changes to cell surface proteins that can easily be applied to other S-layer forming bacteria.}, journal={FRONTIERS IN MICROBIOLOGY}, publisher={Frontiers Media SA}, author={Klotz, Courtney and O'Flaherty, Sarah and Goh, Yong Jun and Barrangou, Rodolphe}, year={2017}, month={Nov} }