Author: Jasenosky, Luke D.; Cadena, Cristhian; Mire, Chad E.; Borisevich, Viktoriya; Haridas, Viraga; Ranjbar, Shahin; Nambu, Aya; Bavari, Sina; Soloveva, Veronica; Sadukhan, Supriya; Cassell, Gail H.; Geisbert, Thomas W.; Hur, Sun; Goldfeld, Anne E.
Title: The FDA-Approved Oral Drug Nitazoxanide Amplifies Host Antiviral Responses and Inhibits Ebola Virus Document date: 2019_8_8
ID: yomg30hg_43
Snippet: To stably co-express dCas9-KRAB and a guide RNA specific to the promoters of the genes encoding i) PKR, ii) RIG-I, or iii) GADD34, a lentiviral vector was constructed that encoded an EFS promoter-driven dCas9-KRAB-2A-E2Crimson-2A-puroR cassette and a U6 promoter for gRNA transcription (see Supp. Fig. 1 ) based on similar approaches our laboratory has used previously (Chow et al., 2014) . Individual gRNA sequences were obtained from the CRISPR des.....
Document: To stably co-express dCas9-KRAB and a guide RNA specific to the promoters of the genes encoding i) PKR, ii) RIG-I, or iii) GADD34, a lentiviral vector was constructed that encoded an EFS promoter-driven dCas9-KRAB-2A-E2Crimson-2A-puroR cassette and a U6 promoter for gRNA transcription (see Supp. Fig. 1 ) based on similar approaches our laboratory has used previously (Chow et al., 2014) . Individual gRNA sequences were obtained from the CRISPR design program at crispr.mit.edu and are shown in Supplemental Figure 1 , along with the sites in the individual gene promoters that they target. As a control, a lentiviral vector encoding a U6 promoter-driven scrambled gRNA that does not match any genomic sequence was also constructed (designated CRISPRcontrol).
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