Human TREX1 Knockout Cell Line-HeLa
Cat.No. : CSC-RT1457
Host Cell: HeLa Target Gene: TREX1
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT1457 |
| Cell Line Information | A stable cell line with a homozygous knockout of human TREX1 using CRISPR/Cas9. |
| Target Gene | TREX1 |
| Host Cell | HeLa |
| Shipping | 10^6 cells/tube |
| Storage | Liquid nitrogen |
| Species | Human |
| Gene ID | 11277 |
| 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. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:4-1:6. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| 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
TREX1, Three-prime Repair Exonuclease 1, is a highly conserved enzyme that is essential in human biology. As a DNA exonuclease, TREX1 plays a key role in maintaining genomic integrity by degrading both single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA). This enzyme is essential to prevent the accumulation of cytoplasmic DNA, thereby avoiding inappropriate activation of immune responses. TREX1 trims nucleotides at the 3' end of DNA, thereby facilitating the removal of abnormal DNA structures, such as those caused by oxidative damage or replication stress. This exonucleolytic activity is essential for resolving stalled replication forks and preventing genomic instability.
Mutations in the TREX1 gene have been associated with various autoimmune and inflammatory diseases. For example, TREX1 deficiency is known to cause Aicardi-Goutier syndrome (AGS), a rare congenital neuroinflammatory disease characterized by encephalopathy and elevated interferon levels. Likewise, TREX1 abnormalities can lead to systemic lupus erythematosus (SLE) and familial lupus pernio due to accumulation of endogenous DNA in the cytosol, triggering chronic activation of the immune system.
Chromothripsis and kataegis are common in cancer and may be caused by telomere crisis, a period of genomic instability during tumorigenesis in which depletion of telomere reserves generates unstable dicentric chromosomes. Here, researchers investigated the mechanisms underlying chromothripsis and kataegis using an in vitro telomere crisis model. They found that TREX1, a cytoplasmic exonuclease that promotes the disassembly of dicentric chromosomes, plays an important role in chromosome fragmentation. In the absence of TREX1, genomic alterations caused by telomere crisis mainly involved break-fusion-bridging cycles and simple genomic rearrangements rather than chromosome fragmentation. Furthermore, the study showed that kataegis observed at chromothriptic breakpoints is the consequence of cytosine deamination by APOBEC3B. These data suggest that chromothripsis and kataegis are caused by a combination of nucleolytic processing by TREX1 and cytosine editing by APOBEC3B.
Of the 14 selected T2p1 postcrisis clones with ≥4 copy number (CN) changes in 1× coverage analysis (Figure 1e), 12 (86%) had either chromothripsis or a chromothripsis-like pattern on 30×WGS (Figure 1b,c). In contrast, among the 14 TREX1 knockout clones with complex events analyzed by 30×WGS, only 3 (21%) showed chromothripsis or chromothripsis-like patterns (Figure 1b,c). Taken together with the low-coverage data, these data suggest that chromothripsis is more common when cells undergo telomere crisis in the presence of TREX1. The patterns of structural variation in the postcrisis TREX1 knockout clones showed that other abnormalities emerge instead of chromothripsis (Figure 1c). Whereas the majority (57%) of CN changes in the T2p1 clones were classified as chromothripsis or chromothripsis-like, TREX1 knockout clones predominantly showed BFB and local jump signatures (Figure 1c,d). Consistently, the number of CN changes per event was lower in TREX1 knockout clones than in T2p1 clones (Figure 1d). These data imply that TREX1 knockout cells resolve DNA bridges formed in telomere crisis through simple structural events rather than chromothripsis.
Figure 1. TREX1 promotes chromothripsis. a, Examples of chromothripsis, chromothripsis-like, BFB and local jump patterns in postcrisis clones derived from T2p1 and TREX1 knockout cells. b, Summary of the number of clones that displayed the types of rearrangements shown in a as determined by 30× WGS of 14 T2p1 and TREX1 knockout postcrisis clones with complex events observed in 1×WGS. c, Summary of the number of chromosomes in postcrisis T2p1 and TREX1 knockout clones examined in b that display the indicated rearrangements. d, Plot of the number of CN changes associated with the complex events indicated in postcrisis T2p1 and TREX1 knockout clones described in b. (Maciejowski, John, et al. 2020)
TREX1, three prime repair exonuclease 1, is an enzyme essential for DNA repair and maintaining genome integrity. The human TREX1 knockout cell line - HeLa can be used for a variety of applications:
Cancer Research: TREX1 deficiency affects cancer cell proliferation, apoptosis, and cellular responses to DNA damage. This knockout cell line allows scientists to dissect the role of TREX1 in tumorigenesis and cancer progression, potentially identifying new therapeutic targets or elucidating mechanisms of resistance to existing treatments.
Autoimmune Disease Research: TREX1 mutations have been associated with autoimmune diseases such as Aicardi-Goutières syndrome and systemic lupus erythematosus. By studying TREX1 knockout HeLa cells, researchers can better understand the pathogenesis of these diseases, including the accumulation of cytoplasmic DNA and activation of innate immune responses.
Viral Infection Mechanisms: TREX1 has been shown to degrade viral cytoplasmic DNA, thereby preventing the immune system from detecting viral DNA. The knockout cell line can be used to study how viruses such as HIV and herpes simplex virus manage to evade host defenses, providing insights that can aid in the development of antiviral therapies.
DNA Repair and Genomic Stability: TREX1 is essential for removing abnormal DNA structures and repairing single-strand DNA breaks. Researchers can use knockout cells to study DNA repair pathways and cellular responses to DNA damage.
Drug Screening and Development: TREX1 knockout cell lines can be used for high-throughput drug screening to identify molecules that specifically affect TREX1-regulated pathways. This could facilitate the development of novel drugs for the treatment of diseases associated with DNA damage and repair, including cancer and autoimmune diseases.
For research use only. Not intended for any clinical use.
