Human IFIT1 Knockout Cell Line-A549
Cat.No. : CSC-RT2121
Host Cell: A549 Target Gene: IFIT1
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2121 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of human IFIT1 using CRISPR/Cas9. |
| Target Gene | IFIT1 |
| Host Cell | A549 |
| Storage | Liquid nirtogen |
| Species | Human |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
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Background
Case Study
Applications
The IFIT1 gene, also known as interferon-induced tetratricopeptide repeat 1 protein, is part of a family of genes that play a key role in the body's immune response to viral infections. The gene is induced by type I interferons, which are cytokines produced in response to viral infections. Interferons activate the expression of several genes that contribute to the cell's antiviral state, of which IFIT1 is a key player. Upon activation, IFIT1 produces a protein that can inhibit viral replication. It does this by binding to viral RNA, thereby preventing the virus from hijacking the host cell's machinery to produce its own proteins.
The IFIT1 protein is characterized by the presence of multiple tetratricopeptide repeats (TPRs), which are structural motifs involved in protein-protein interactions. These TPRs facilitate IFIT1 binding to other cellular proteins and viral components, thereby mediating its antiviral effects. For example, IFIT1 can recognize and bind to the 5'-triphosphate end of viral RNAs, a molecular feature that distinguishes them from mature host RNAs. This binding inhibits the translation of the viral genome into proteins, effectively delaying the viral life cycle. In addition to its antiviral function, IFIT1 is also involved in regulating the host's immune response. It can interact with other components of the immune signaling pathway to enhance the body's defenses. Studies have also shown that IFIT1 plays an important role in the pathogenesis of inflammatory responses and autoimmune diseases.
Members of the interferon-inducible protein with tetratricopeptide repeats (IFIT) family of proteins are highly expressed in response to type I IFN signaling and have been shown to interfere with viral protein synthesis and modulate innate immune signaling. Specifically, IFIT1 has been reported by multiple independent research groups to interfere with viral replication by interacting with unmethylated viral mRNA in the absence of viral 2-O'-MTase activity, thereby discriminating between viral and host mRNAs.
Here, to test whether the hypersensitivity of SARS-CoV-2 to IFNs lacking functional Nsp16 is attributable to the activity of IFIT1, ACE2-expressing A549 control cells and IFIT1 knockout (KO) cells were preincubated with increasing amounts of type I IFN 18 h before infection with wt or Nsp16mut virus (Figure 1A). By RT-qPCR, increasing amounts of type I IFN reduced SARS-CoV-2 wt replication in A549 control cells and IFIT1 KO cells at 3 dpi. However, in the case of Nsp16mut infection, the researchers found that viral replication was very strongly blocked after treatment with increasing doses of type I IFN (Figure 1A), which was inversely correlated with increased IFIT1 expression levels (Figure 1B). Surprisingly, the dose-dependent inhibitory effect of type I IFN on Nsp16mut was absent in cells lacking the restriction factor IFIT1. Similarly, the researchers generated Calu-3 IFIT1 KO cells as well as control KO cells by lentiviral transduction followed by antibiotic selection (Figure 1C and D). Although the polyclonal cell lines showed residual IFIT1 expression, type I IFN-mediated blockade of Nsp16mut replication was almost completely abolished in both IFIT1 KO cell lines (Figure 1C). The results indicate that IFIT1 is the major restriction factor that blocks SARS-CoV-2 replication in response to type I IFN but is counteracted by the viral 2′-O-methyltransferase Nsp16.
Figure 1. The high sensitivity of SARS–CoV‐2 Nsp16mut to type I IFN is mediated by IFIT1. (Russ A, et al., 2022)
The Human IFIT1 Knockout Cell Line-A549 is a valuable model for a variety of research applications, particularly in the fields of virology, immunology, and oncology. Here are some key applications:
Viral Infection Studies: The Human IFIT1 Knockout Cell Line-A549 is essential for studying the role of interferon-induced proteins in antiviral responses.
Drug Discovery and Development: This cell line is valuable in screening potential antiviral drugs. By knocking out IFIT1, scientists can identify compounds that can compensate for the loss of this protein, thereby aiding the development of new therapeutics for viral infections.
Immune Response Analysis: Researchers can assess how IFIT1 loss affects the expression of other genes and proteins involved in the immune response, providing insight into the complex immune signaling network.
Cancer Research: IFIT1 has been implicated in the regulation of cell growth and apoptosis. The use of the knockout cell line in A549 lung cancer cells has helped to understand its role in tumorigenesis, potentially leading to new therapeutic targets for lung cancer treatment.
Interferon Pathway Studies: This cell line can serve as a model for studying the broader interferon signaling pathway. By comparing the responses of knockout and wild-type cells, the researchers could dissect the specific contribution of IFIT1 within the interferon-stimulated gene network.
For research use only. Not intended for any clinical use.
