{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T23:47:29Z","timestamp":1790207249999,"version":"4.1.0"},"reference-count":49,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2015,5,12]],"date-time":"2015-05-12T00:00:00Z","timestamp":1431388800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Department of Defense Chemical and Biological Defense program through the Defense Threat Reduction Agency (DTRA).","award":["HDTRA1\u201310-C-0063"],"award-info":[{"award-number":["HDTRA1\u201310-C-0063"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2015,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>We used a genomic library of mutant murine embryonic stem cells (ESCs) and report the methodology required to simultaneously culture, differentiate, and screen more than 3,200 heterozygous mutant clones to identify host-based genes involved in both sensitivity and resistance to rabies virus infection. Established neuronal differentiation protocols were miniaturized such that many clones could be handled simultaneously, and molecular markers were used to show that the resultant cultures were pan-neuronal. Next, we used a green fluorescent protein (GFP) labeled rabies virus to develop, validate, and implement one of the first host-based, high-content, high-throughput screens for rabies virus. Undifferentiated cell and neuron cultures were infected with GFP-rabies and live imaging was used to evaluate GFP intensity at time points corresponding to initial infection\/uptake and early and late replication. Furthermore, supernatants were used to evaluate viral shedding potential. After repeated testing, 63 genes involved in either sensitivity or resistance to rabies infection were identified. To further explore hits, we used a completely independent system (siRNA) to show that reduction in target gene expression leads to the observed phenotype. We validated the immune modulatory gene Unc13d and the dynein adapter gene Bbs4 by treating wild-type ESCs and primary neurons with siRNA; treated cultures were resistant to rabies infection\/replication. Overall, the potential of such in vitro functional genomics screens in stem cells adds additional value to other libraries of stem cells. This technique is applicable to any bacterial or virus interactome and any cell or tissue types that can be differentiated from ESCs. Stem Cells \u00a02015;33:2509\u20132522<\/jats:p>","DOI":"10.1002\/stem.1983","type":"journal-article","created":{"date-parts":[[2015,3,5]],"date-time":"2015-03-05T22:54:34Z","timestamp":1425596074000},"page":"2509-2522","source":"Crossref","is-referenced-by-count":2,"title":["High-Throughput Differentiation and Screening of a Library of Mutant Stem Cell Clones Defines New Host-Based Genes Involved in Rabies Virus Infection"],"prefix":"10.1093","volume":"33","author":[{"given":"Deeann","family":"Wallis","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Biophysics Texas A&M University, College Station, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kimberly","family":"Loesch","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Biophysics Texas A&M University, College Station, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stacy","family":"Galaviz","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Biophysics Texas A&M University, College Station, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingan","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Biophysics Texas A&M University, College Station, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"DeJesus","sequence":"additional","affiliation":[{"name":"Department of Computer Science and Engineering Texas A&M University, College Station, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Ioerger","sequence":"additional","affiliation":[{"name":"Department of Computer Science and Engineering Texas A&M University, College Station, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James C.","family":"Sacchettini","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Biophysics Texas A&M University, College Station, Texas, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2015,5,12]]},"reference":[{"key":"2022011112055312000_stem1983-bib-0001","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.cell.2006.12.018","article-title":"A mouse for all reasons","volume":"128","author":"Consortium","year":"2007","journal-title":"Cell"},{"key":"2022011112055312000_stem1983-bib-0002","doi-asserted-by":"crossref","first-page":"1670","DOI":"10.1101\/gr.078352.108","article-title":"Large-scale gene trapping in C57BL\/6N mouse embryonic stem cells","volume":"18","author":"Hansen","year":"2008","journal-title":"Genome Res"},{"key":"2022011112055312000_stem1983-bib-0003","doi-asserted-by":"crossref","first-page":"bar028","DOI":"10.1093\/database\/bar028","article-title":"BioMart as an integration solution for the International Knockout Mouse Consortium","volume":"2011","author":"Oakley","year":"2011","journal-title":"Database (Oxford)"},{"key":"2022011112055312000_stem1983-bib-0004","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1016\/j.ymeth.2010.12.022","article-title":"Genetic screens using the piggyBac transposon","volume":"53","author":"Chew","year":"2011","journal-title":"Methods"},{"key":"2022011112055312000_stem1983-bib-0005","doi-asserted-by":"crossref","first-page":"R48","DOI":"10.1186\/gb-2007-8-4-r48","article-title":"A recessive genetic screen for host factors required for retroviral infection in a library of insertionally mutated Blm-deficient embryonic stem cells","volume":"8","author":"Wang","year":"2007","journal-title":"Genome Biol"},{"key":"2022011112055312000_stem1983-bib-0006","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/S0076-6879(10)77012-9","article-title":"Genome-wide forward genetic screens in mouse ES cells","volume":"477","author":"Li","year":"2010","journal-title":"Methods Enzymol"},{"key":"2022011112055312000_stem1983-bib-0007","doi-asserted-by":"crossref","first-page":"891","DOI":"10.1038\/nature02653","article-title":"Mismatch repair genes identified using genetic screens in Blm-deficient embryonic stem cells","volume":"429","author":"Guo","year":"2004","journal-title":"Nature"},{"key":"2022011112055312000_stem1983-bib-0008","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1038\/nrmicro2260","article-title":"The cell biology of rabies virus: Using stealth to reach the brain","volume":"8","author":"Schnell","year":"2010","journal-title":"Nat Rev Microbiol"},{"key":"2022011112055312000_stem1983-bib-0009","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/S0065-3527(08)00005-5","article-title":"Lyssaviruses: Current trends","volume":"71","author":"Nadin-Davis","year":"2008","journal-title":"Adv Virus Res"},{"key":"2022011112055312000_stem1983-bib-0010","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/B978-0-12-387040-7.00006-8","article-title":"Interferon in rabies virus infection","volume":"79","author":"Rieder","year":"2011","journal-title":"Adv Virus Res"},{"key":"2022011112055312000_stem1983-bib-0011","doi-asserted-by":"crossref","first-page":"7229","DOI":"10.1073\/pnas.0701397104","article-title":"The dynein light chain 8 binding motif of rabies virus phosphoprotein promotes efficient viral transcription","volume":"104","author":"Tan","year":"2007","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022011112055312000_stem1983-bib-0012","doi-asserted-by":"crossref","first-page":"e33561","DOI":"10.1371\/journal.pone.0033561","article-title":"The 3' untranslated region of the rabies virus glycoprotein mRNA specifically interacts with cellular PCBP2 protein and promotes transcript stability","volume":"7","author":"Palusa","year":"2012","journal-title":"PLoS One"},{"key":"2022011112055312000_stem1983-bib-0013","doi-asserted-by":"crossref","first-page":"2758","DOI":"10.1073\/pnas.051630298","article-title":"Effect of rabies virus infection on gene expression in mouse brain","volume":"98","author":"Prosniak","year":"2001","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022011112055312000_stem1983-bib-0014","doi-asserted-by":"crossref","first-page":"12554","DOI":"10.1128\/JVI.79.19.12554-12565.2005","article-title":"Attenuated rabies virus activates, while pathogenic rabies virus evades, the host innate immune responses in the central nervous system","volume":"79","author":"Wang","year":"2005","journal-title":"J Virol"},{"key":"2022011112055312000_stem1983-bib-0015","doi-asserted-by":"crossref","first-page":"12893","DOI":"10.1128\/JVI.79.20.12893-12904.2005","article-title":"Virus infection switches TLR-3-positive human neurons to become strong producers of beta interferon","volume":"79","author":"Prehaud","year":"2005","journal-title":"J Virol"},{"key":"2022011112055312000_stem1983-bib-0016","doi-asserted-by":"crossref","first-page":"951","DOI":"10.1111\/j.1348-0421.2006.tb03871.x","article-title":"Screening of upregulated genes in suckling mouse central nervous system during the disease stage of rabies virus infection","volume":"50","author":"Ubol","year":"2006","journal-title":"Microbiol Immunol"},{"key":"2022011112055312000_stem1983-bib-0017","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1080\/13550280601178226","article-title":"Proteomic profiling reveals that rabies virus infection results in differential expression of host proteins involved in ion homeostasis and synaptic physiology in the central nervous system","volume":"13","author":"Dhingra","year":"2007","journal-title":"J Neurovirol"},{"key":"2022011112055312000_stem1983-bib-0018","doi-asserted-by":"crossref","first-page":"2399","DOI":"10.1002\/pmic.200701007","article-title":"Proteomics analysis of BHK-21 cells infected with a fixed strain of rabies virus","volume":"9","author":"Zandi","year":"2009","journal-title":"Proteomics"},{"key":"2022011112055312000_stem1983-bib-0019","doi-asserted-by":"crossref","first-page":"5493","DOI":"10.1007\/s00253-013-4939-1","article-title":"Proteome analysis of virus-host cell interaction: Rabies virus replication in Vero cells in two different media","volume":"97","author":"Kluge","year":"2013","journal-title":"Appl Microbiol Biotechnol"},{"key":"2022011112055312000_stem1983-bib-0020","doi-asserted-by":"crossref","first-page":"1640","DOI":"10.1128\/JVI.02602-14","article-title":"The focal adhesion kinase is involved in rabies virus infection through its interaction with the viral phosphoprotein P","volume":"89","author":"Fouquet","year":"2015","journal-title":"J Virol"},{"key":"2022011112055312000_stem1983-bib-0021","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1016\/j.coviro.2012.09.003","article-title":"Virus-host interactomes\u2014Antiviral drug discovery","volume":"2","author":"Ma-Lauer","year":"2012","journal-title":"Curr Opin Virol"},{"key":"2022011112055312000_stem1983-bib-0022","doi-asserted-by":"crossref","first-page":"E861","DOI":"10.1073\/pnas.1210198110","article-title":"Host-rabies virus protein-protein interactions as druggable antiviral targets","volume":"110","author":"Lingappa","year":"2013","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022011112055312000_stem1983-bib-0023","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/0925-4773(95)00446-8","article-title":"Differentiation of pluripotent embryonic stem cells into the neuronal lineage in vitro gives rise to mature inhibitory and excitatory neurons","volume":"53","author":"Strubing","year":"1995","journal-title":"Mech Dev"},{"key":"2022011112055312000_stem1983-bib-0024","doi-asserted-by":"crossref","first-page":"17209","DOI":"10.1074\/jbc.M109.098814","article-title":"High-throughput screening in embryonic stem cell-derived neurons identifies potentiators of alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionate-type glutamate receptors","volume":"285","author":"McNeish","year":"2010","journal-title":"J Biol Chem"},{"key":"2022011112055312000_stem1983-bib-0025","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1016\/j.cell.2005.08.012","article-title":"Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-kappaB and IRF 3","volume":"122","author":"Seth","year":"2005","journal-title":"Cell"},{"key":"2022011112055312000_stem1983-bib-0026","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1038\/nbt1186","article-title":"Statistical practice in high-throughput screening data analysis","volume":"24","author":"Malo","year":"2006","journal-title":"Nat Biotechnol"},{"key":"2022011112055312000_stem1983-bib-0027","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1038\/nn1301","article-title":"Differentiation of mouse embryonic stem cells into a defined neuronal lineage","volume":"7","author":"Bibel","year":"2004","journal-title":"Nat Neurosci"},{"key":"2022011112055312000_stem1983-bib-0028","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/j.bbrc.2010.12.132","article-title":"Embryonic stem cell-derived neurons are a novel, highly sensitive tissue culture platform for botulinum research","volume":"405","author":"McNutt","year":"2011","journal-title":"Biochem Biophys Res Commun"},{"key":"2022011112055312000_stem1983-bib-0029","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.jviromet.2004.12.003","article-title":"A novel rapid fluorescent focus inhibition test for rabies virus using a recombinant rabies virus visualizing a green fluorescent protein","volume":"125","author":"Khawplod","year":"2005","journal-title":"J Virol Methods"},{"key":"2022011112055312000_stem1983-bib-0030","doi-asserted-by":"crossref","first-page":"9405","DOI":"10.1073\/pnas.0403060101","article-title":"Rabies virus nucleoprotein as a carrier for foreign antigens","volume":"101","author":"Koser","year":"2004","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022011112055312000_stem1983-bib-0031","doi-asserted-by":"crossref","first-page":"e1003941","DOI":"10.1371\/journal.ppat.1003941","article-title":"CIB1 synergizes with EphrinA2 to regulate Kaposi's sarcoma-associated herpesvirus macropinocytic entry in human microvascular dermal endothelial cells","volume":"10","author":"Bandyopadhyay","year":"2014","journal-title":"PLoS Pathog"},{"key":"2022011112055312000_stem1983-bib-0032","doi-asserted-by":"crossref","first-page":"4401","DOI":"10.1242\/jcs.01300","article-title":"A role for the PDZ-binding domain of the coxsackie B virus and adenovirus receptor (CAR) in cell adhesion and growth","volume":"117","author":"Excoffon","year":"2004","journal-title":"J Cell Sci"},{"key":"2022011112055312000_stem1983-bib-0033","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1002\/(SICI)1097-0177(199806)212:2<304::AID-AJA15>3.0.CO;2-3","article-title":"Aquarius, a novel gene isolated by gene trapping with an RNA-dependent RNA polymerase motif","volume":"212","author":"Sam","year":"1998","journal-title":"Dev Dyn"},{"key":"2022011112055312000_stem1983-bib-0034","doi-asserted-by":"crossref","first-page":"4551","DOI":"10.1128\/JVI.01366-06","article-title":"Participation of rab5, an early endosome protein, in hepatitis C virus RNA replication machinery","volume":"81","author":"Stone","year":"2007","journal-title":"J Virol"},{"key":"2022011112055312000_stem1983-bib-0035","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1038\/36599","article-title":"DAP kinase links the control of apoptosis to metastasis","volume":"390","author":"Inbal","year":"1997","journal-title":"Nature"},{"key":"2022011112055312000_stem1983-bib-0036","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.virol.2011.01.032","article-title":"Latent membrane protein 1 of Epstein-Barr virus regulates death-associated protein kinase 1 in lymphoblastoid cell line","volume":"413","author":"Lee","year":"2011","journal-title":"Virology"},{"key":"2022011112055312000_stem1983-bib-0037","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/j.virol.2006.01.047","article-title":"Interactions of the cellular CCAAT displacement protein and human papillomavirus E2 protein with the viral origin of replication can regulate DNA replication","volume":"350","author":"Narahari","year":"2006","journal-title":"Virology"},{"key":"2022011112055312000_stem1983-bib-0038","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1016\/j.virol.2008.05.024","article-title":"Repression of human cytomegalovirus major immediate early gene expression by the cellular transcription factor CCAAT displacement protein","volume":"378","author":"Stern","year":"2008","journal-title":"Virology"},{"key":"2022011112055312000_stem1983-bib-0039","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.semcancer.2009.03.002","article-title":"Cellular transformation by Simian Virus 40 and Murine Polyoma Virus T antigens","volume":"19","author":"Cheng","year":"2009","journal-title":"Semin Cancer Biol"},{"key":"2022011112055312000_stem1983-bib-0040","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1016\/j.neulet.2005.03.044","article-title":"E2F4 expression patterns in SIV encephalitis","volume":"382","author":"Morgan","year":"2005","journal-title":"Neurosci Lett"},{"key":"2022011112055312000_stem1983-bib-0041","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/S1097-2765(00)00030-7","article-title":"Loss of E2F4 activity leads to abnormal development of multiple cellular lineages","volume":"6","author":"Rempel","year":"2000","journal-title":"Mol Cell"},{"key":"2022011112055312000_stem1983-bib-0042","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1016\/S0092-8674(03)00855-9","article-title":"Munc13-4 is essential for cytolytic granules fusion and is mutated in a form of familial hemophagocytic lymphohistiocytosis (FHL3)","volume":"115","author":"Feldmann","year":"2003","journal-title":"Cell"},{"key":"2022011112055312000_stem1983-bib-0043","doi-asserted-by":"crossref","first-page":"489","DOI":"10.3390\/toxins3050489","article-title":"Alpha-latrotoxin rescues SNAP-25 from BoNT\/A-mediated proteolysis in embryonic stem cell-derived neurons","volume":"3","author":"Mesngon","year":"2011","journal-title":"Toxins (Basel)"},{"key":"2022011112055312000_stem1983-bib-0044","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1084\/jem.20062447","article-title":"Jinx, an MCMV susceptibility phenotype caused by disruption of Unc13d: A mouse model of type 3 familial hemophagocytic lymphohistiocytosis","volume":"204","author":"Crozat","year":"2007","journal-title":"J Exp Med"},{"key":"2022011112055312000_stem1983-bib-0045","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1038\/ng1352","article-title":"The Bardet-Biedl protein BBS4 targets cargo to the pericentriolar region and is required for microtubule anchoring and cell cycle progression","volume":"36","author":"Kim","year":"2004","journal-title":"Nat Genet"},{"key":"2022011112055312000_stem1983-bib-0046","doi-asserted-by":"crossref","first-page":"10212","DOI":"10.1128\/JVI.74.21.10212-10216.2000","article-title":"Interaction of the rabies virus P protein with the LC8 dynein light chain","volume":"74","author":"Raux","year":"2000","journal-title":"J Virol"},{"key":"2022011112055312000_stem1983-bib-0047","doi-asserted-by":"crossref","first-page":"10217","DOI":"10.1128\/JVI.74.21.10217-10222.2000","article-title":"Cytoplasmic dynein LC8 interacts with lyssavirus phosphoprotein","volume":"74","author":"Jacob","year":"2000","journal-title":"J Virol"},{"key":"2022011112055312000_stem1983-bib-0048","doi-asserted-by":"crossref","first-page":"3204","DOI":"10.1091\/mbc.e07-01-0030","article-title":"Dynein light chain association sequences can facilitate nuclear protein import","volume":"18","author":"Moseley","year":"2007","journal-title":"Mol Biol Cell"},{"key":"2022011112055312000_stem1983-bib-0049","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/j.cell.2008.10.007","article-title":"Knockdown screens to knockout HIV-1","volume":"135","author":"Goff","year":"2008","journal-title":"Cell"}],"container-title":["Stem Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fstem.1983","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fstem.1983","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/stem.1983","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/stem.1983","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/academic.oup.com\/stmcls\/article-pdf\/33\/8\/2509\/42018423\/stmcls_33_8_2509.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/stmcls\/article-pdf\/33\/8\/2509\/42018423\/stmcls_33_8_2509.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,1,19]],"date-time":"2022-01-19T02:55:18Z","timestamp":1642560918000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/stmcls\/article\/33\/8\/2509-2522\/6432092"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,5,12]]},"references-count":49,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2015,5,12]]},"published-print":{"date-parts":[[2015,8,1]]}},"URL":"https:\/\/doi.org\/10.1002\/stem.1983","relation":{},"ISSN":["1066-5099","1549-4918"],"issn-type":[{"value":"1066-5099","type":"print"},{"value":"1549-4918","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2015,8,1]]},"published":{"date-parts":[[2015,5,12]]}}}