Ifit1

Manufactured by Thermo Fisher Scientific

The Ifit1 is a laboratory equipment designed for nucleic acid detection and analysis. It functions as a sensor that recognizes and binds to specific RNA sequences, enabling the identification and quantification of target molecules.

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Lab products found in correlation

4 protocols using ifit1

Gene Expression Profiling Assay

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JMJD1a, JMJD1b, JMJD2a, JMJD2b, JMJD2c, JMJD2d, LSD1, PHF8, Ccl5, Mx2, IRF7, Ifit1, and Ifit3 TaqMan probes and JMJD2d siRNAs were obtained from Applied Biosystems. All other primers were designed and obtained from Sigma and are summarized in Supplemental Tables 2, 3. All forward and reverse primers were mixed to a final concentration of 50 μM.

Chandwani R., Fang T.C., Dewell S, & Tarakhovsky A. (2023). Control of enhancer and promoter activation in the type I interferon response by the histone demethylase Kdm4d/JMJD2d. Frontiers in Immunology, 14, 1146699.

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Quantitative Gene Expression Analysis

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Total RNA was extracted using an RNeasy kit (Qiagen, Valencia, CA). Messenger RNA (mRNA) expression was measured by quantitative real-time PCR (7300 Real-Time PCR System; Applied Biosystems, Foster City, CA) with TaqMan Gene Expression Assays (Applied Biosystems), according to the manufacturer's instructions. The expression levels of the following genes were measured: NIS (Hs00166567_m1), STAT1 (Hs01013996_m1), STAT2 (Hs01013115_g1), IRF1 (Hs00971965_m1), and IFIT1 (Hs03027069_s1) (Applied Biosystems). Expression was normalized for each gene to the housekeeping gene betaactin (ACTB, Hs01060665_g1; Applied Biosystems). Assays were performed in triplicate.

Shiozaki A., Ariyoshi Y., Iitaka D., Kosuga T., Shimizu H., Kudou M., Konishi T., Shoda K., Arita T., Konishi H., Komatsu S., Kubota T., Fujiwara H., Okamoto K., Kishimoto M., Konishi E., Marunaka Y., Ichikawa D, & Otsuji E. (2019). Functional analysis and clinical significance of sodium iodide symporter expression in gastric cancer. Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association, 22(3).

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Quantifying Radiation-Induced Gene Expression

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Real-time quantitative RT-PCR (qRT-PCR) was performed for selected genes using Taqman chemistry and the ABI 7900 Real Time PCR System. The Globin cleared purified RNA from 5 control and irradiated animals was used for cDNA synthesis using the High-Capacity cDNA Archive Kit (Life Technologies). Gene expression assays (primer/probe sets) were purchased from Thermo Fisher for the following genes: Ifit3 (Mm00366278_m1), Ifit1 (Mm00365614_m1), Cdkn1a (Mm04205640_g1), Aen (Mm00471554_m1), Gstt1 (Mm00492506_m1), Il18r1 (Mm00515178_m1), Ccng1 (Mm00438084_m1), Alas1 (Mm001235914_m1), Acsbg1 (Mm00547366_m1), Cyp2b6 (Mn00456591_m1), Actb (Mm00607939_s1). The ΔΔCT method was used to calculate expression relative to controls, using normalization to Actb expression. Each reaction was run in triplicate with 5 control and 5 irradiated samples, and means were compared using an unpaired t-test.

Mukherjee S., Laiakis E.C., Fornace AJ J.r, & Amundson S.A. (2019). Impact of inflammatory signaling on radiation biodosimetry: mouse model of inflammatory bowel disease. BMC Genomics, 20, 329.

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Quantitative PCR analysis of human liver

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Whole liver RNA was isolated using RNeasy mini kit (Qiagen, Hilden, Germany) including DNAse treatment according to manufacturer’s protocol starting with homogenization of liver tissue in RLT buffer. cDNA was generated by using an iScript cDNA synthesis kit (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s protocol. Human specific gene expression was measured using Taqman primer/probe quantitative PCR, in TaqMan® Gene Expression Master Mix (Thermo Fisher Scientific, Waltham, MA, USA). Primer/probe combinations were purchased from Thermo Fisher Scientific; CXCL10 (Hs01124251_g1), CXCL9 (Hs00171065_m1), DDX58 (Hs01061436_m1), GAPDH (Hs00266705_g1), IFIT1 (Hs01911452_s1), ISG15 (Hs01921425_s1), IFNA1 (Hs00855471_g1), IFNA4 (Hs01681284_sh), IFNB1 (Hs01077958_s1), MX1 (Hs00895608_m1), OAS1 (Hs00973637_m1), RSAD2 (Hs00369813_m1), STAT1 (Hs01013996_m1), TLR3 (Hs01551078_m1). Expression of target genes was normalized to the expression of GAPDH using the formula 2^−ΔCt, ΔCt = Ct^target−Ct^GADPH. cDNA from non-chimeric mouse livers was used as control to test cross-reactivity of housekeeping and target genes. Due to the difference in hepatocyte donor baseline expression levels of examined genes (Suppl. Fig. 1), fold changes of transcripts were calculated to those of non-infected humanized livers from mice transplanted with the identical hepatocyte donor.

van de Garde M.D., Pas S.D., van Oord G.W., Gama L., Choi Y., de Man R.A., Boonstra A, & Vanwolleghem T. (2017). Interferon-alpha treatment rapidly clears Hepatitis E virus infection in humanized mice. Scientific Reports, 7, 8267.

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