IRF1 Knockout RBE Cell Pool
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Product ID： LM01085125347
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隐藏域元素占位
- 产品描述
- 细胞复苏
- 细胞传代
- 细胞冻存
- 抗体验证结果
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- Brand: ELEM粒曼
- Commodity name: IRF1 Knockout RBE Cell Pool
- Commodity ID: LM01085125347
- Gene Symbol: IRF1
- Ensembl ID: ENSG00000125347
- Uniprot ID: P10914
- 宿主细胞 / 类型: RBE/human hepatobiliary carcinoma cells
- NCBI Gene ID: 3659
- 规格: 1×10^6 cells/frozen vial
- 生长培养基: 89% 1640 + 10% FBS + 1% double antibody
- 筛选标记: N/A
- 生长特性: Adherent cells, epithelial-like
- 培养条件: Incubator at 37°C with 5% CO2; passage at 1/2 to 1/4 confluence.
- 倍增时间: ~24-36 hours
- 参考换液频率: 2–3 times per week
- 支原体检测结果: Negative
- 敲除效率（Sanger测序）: 97%
- 蛋白质组验证结果: N/A
- 抗体货号: Adding...
- 目标基因介绍: Transcriptional regulator which displays a remarkable functional diversity in the regulation of cellular responses (PubMed:15226432, PubMed:15509808, PubMed:17516545, PubMed:17942705, PubMed:18497060, PubMed:19404407, PubMed:19851330, PubMed:22367195, PubMed:32385160). Regulates transcription of IFN and IFN-inducible genes, host response to viral and bacterial infections, regulation of many genes expressed during hematopoiesis, inflammation, immune responses and cell proliferation and differentiation, regulation of the cell cycle and induction of growth arrest and programmed cell death following DNA damage (PubMed:15226432, PubMed:15509808, PubMed:17516545, PubMed:17942705, PubMed:18497060, PubMed:19404407, PubMed:19851330, PubMed:22367195). Stimulates both innate and acquired immune responses through the activation of specific target genes and can act as a transcriptional activator and repressor regulating target genes by binding to an interferon-stimulated response element (ISRE) in their promoters (PubMed:15226432, PubMed:15509808, PubMed:17516545, PubMed:17942705, PubMed:18497060, PubMed:19404407, PubMed:19851330, PubMed:21389130, PubMed:22367195). Competes with the transcriptional repressor ZBED2 for binding to a common consensus sequence in gene promoters (PubMed:32385160). Its target genes for transcriptional activation activity include: genes involved in anti-viral response, such as IFN-alpha/beta, DDX58/RIG-I, TNFSF10/TRAIL, ZBP1, OAS1/2, PIAS1/GBP, EIF2AK2/PKR and RSAD2/viperin; antibacterial response, such as NOS2/INOS; anti-proliferative response, such as p53/TP53, LOX and CDKN1A; apoptosis, such as BBC3/PUMA, CASP1, CASP7 and CASP8; immune response, such as IL7, IL12A/B and IL15, PTGS2/COX2 and CYBB; DNA damage responses and DNA repair, such as POLQ/POLH; MHC class I expression, such as TAP1, PSMB9/LMP2, PSME1/PA28A, PSME2/PA28B and B2M and MHC class II expression, such as CIITA; metabolic enzymes, such as ACOD1/IRG1 (PubMed:15226432, PubMed:15509808, PubMed:17516545, PubMed:17942705, PubMed:18497060, PubMed:19404407, PubMed:19851330, PubMed:22367195). Represses genes involved in anti-proliferative response, such as BIRC5/survivin, CCNB1, CCNE1, CDK1, CDK2 and CDK4 and in immune response, such as FOXP3, IL4, ANXA2 and TLR4 (PubMed:18641303, PubMed:22200613). Stimulates p53/TP53-dependent transcription through enhanced recruitment of EP300 leading to increased acetylation of p53/TP53 (PubMed:15509808, PubMed:18084608). Plays an important role in immune response directly affecting NK maturation and activity, macrophage production of IL12, Th1 development and maturation of CD8+ T-cells (PubMed:11244049, PubMed:11846971, PubMed:11846974, PubMed:16932750). Also implicated in the differentiation and maturation of dendritic cells and in the suppression of regulatory T (Treg) cells development (PubMed:11244049, PubMed:11846971, PubMed:11846974, PubMed:16932750). Acts as a tumor suppressor and plays a role not only in antagonism of tumor cell growth but also in stimulating an immune response against tumor cells (PubMed:20049431).
- 细胞开发路径: A stable KO cell pool was generated using the CRISPR‑RNP approach; Sanger sequencing revealed a knockout efficiency of 97% in the cell pool.
- 应用: A gene knockout cell pool with high knockout efficiency (KO Cell Pool) is particularly well suited for preliminary functional analyses, the development of complex disease models, precision drug screening, and broad‑scale gene discovery studies. The KO pool can be directly applied to a variety of assays and analyses without the need for time‑consuming single‑clone selection, significantly enhancing experimental throughput.
Key words:- IRF1
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01. Preheat the complete culture medium in a 37°C water bath.
02. Thaw the cryovial in a 37°C water bath for 1–2 minutes.
03. Transfer the cryovials into a biosafety cabinet and wipe their surfaces with 70% ethanol.
04. Unscrew the cap of the cryovial and gently transfer the cell suspension into a sterile centrifuge tube containing 9 mL of complete culture medium.
05. Centrifuge at room temperature at 125g for 5–7 minutes, then discard the supernatant.
06. Resuspend the cell pellet in 5 mL of complete culture medium, and transfer the cell suspension to a T25 culture flask.
07. Transfer the cells to a 37°C incubator with 5% CO2 for culture.
08. Recommended passage ratio: 1/2 to 1/4; cells reach confluence in 2–3 days. -
01. When the cell confluence in the culture flask reaches 80%–90% or higher, cells can be passaged.
02. Remove the culture medium, PBS, and trypsin (0.25% Trypsin‑EDTA, Gibco 25200‑056) from the 4°C refrigerator, place them in a 37°C water bath, and once their temperature approaches 37°C, take them out. Spray the exterior of the bottles with 75% ethanol, then transfer them to a biosafety cabinet.03. Remove the culture flask to be passaged from the incubator, spray the exterior of the flask with 75% ethanol, and place it inside a biosafety cabinet.
04. To avoid dispersing the cells, rinse the cells with PBS along the inner wall of the culture flask; discard the wash solution, then add 2 mL to a T25 flask.
05. Add the appropriate volume of trypsin (1.5 mL for a T75 flask, 0.5 mL for a T25 flask), and gently swirl the flask to ensure the enzyme evenly covers the cell‑covered bottom. Adjust the volume as needed based on the specific situation. After approximately 1–2 minutes, when most cells have detached, add an equal volume of complete culture medium to stop the digestion, then gently pipette up and down with a 5‑mL pipette until all cells are fully detached.
06. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 300 × g for 5 minutes, and discard the supernatant.
07. Transfer 5 mL of complete culture medium to resuspend the cells, adjust the seeding ratio as needed, and replenish the culture flask with complete medium—add up to 13–15 mL for a T75 flask and 5 mL for a T25 flask—then supplement with 1% double antibiotic.
08. After capping the bottle and tightening the lid, gently shake the vial to ensure uniform mixing of the cells, then place it in a 37°C incubator with 5% CO2. -
01. Prepare the cryopreservation solution and pre-cool it in advance.
02. Ensure that the cells to be cryopreserved meet the cryopreservation requirements, and verify the following conditions under a microscope: healthy appearance and morphological characteristics, being in the late logarithmic growth phase, and showing no signs of contamination or senescence.
03. Digest and centrifuge the cells (refer to the subculture protocol for specific steps).
04. Resuspend the cells in cryopreservation medium at 1 mL per tube, gently pipette up and down to ensure uniformity, then aliquot into cryovials.
05. Place the cells in a programmed cooling box and freeze them in a −80°C freezer.
06. Subsequently, the cells are transferred to a liquid nitrogen tank for long-term storage. - In antibody validation
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Classification： Gene Knockout Cell Pool（KO Pool）
Cell Line Information
Gene Symbol
IRF1
NCBI Gene ID
3659
Ensembl ID
ENSG00000125347
Uniprot ID
P10914
Screening marker
N/A
Host cell/type
RBE/human hepatobiliary carcinoma cells
Specifications
1×10^6 cells/frozen vial
Growth Medium
89% 1640 + 10% FBS + 1% double antibody
growth characteristics
Adherent cells, epithelial-like
culture condition
Incubator at 37°C with 5% CO2; passage at 1/2 to 1/4 confluence.
doubling time
~24-36 hours
Reference fluid change frequency
2–3 times per week
Mycoplasma test results
Negative
Knock-out validation
Knockout efficiency (Sanger sequencing)
97%
Proteome Validation Results
N/A
Antibody number
Adding...
Antibody validation results
Cell Line Description
Introduction of target gene
Transcriptional regulator which displays a remarkable functional diversity in the regulation of cellular responses (PubMed:15226432, PubMed:15509808, PubMed:17516545, PubMed:17942705, PubMed:18497060, PubMed:19404407, PubMed:19851330, PubMed:22367195, PubMed:32385160). Regulates transcription of IFN and IFN-inducible genes, host response to viral and bacterial infections, regulation of many genes expressed during hematopoiesis, inflammation, immune responses and cell proliferation and differentiation, regulation of the cell cycle and induction of growth arrest and programmed cell death following DNA damage (PubMed:15226432, PubMed:15509808, PubMed:17516545, PubMed:17942705, PubMed:18497060, PubMed:19404407, PubMed:19851330, PubMed:22367195). Stimulates both innate and acquired immune responses through the activation of specific target genes and can act as a transcriptional activator and repressor regulating target genes by binding to an interferon-stimulated response element (ISRE) in their promoters (PubMed:15226432, PubMed:15509808, PubMed:17516545, PubMed:17942705, PubMed:18497060, PubMed:19404407, PubMed:19851330, PubMed:21389130, PubMed:22367195). Competes with the transcriptional repressor ZBED2 for binding to a common consensus sequence in gene promoters (PubMed:32385160). Its target genes for transcriptional activation activity include: genes involved in anti-viral response, such as IFN-alpha/beta, DDX58/RIG-I, TNFSF10/TRAIL, ZBP1, OAS1/2, PIAS1/GBP, EIF2AK2/PKR and RSAD2/viperin; antibacterial response, such as NOS2/INOS; anti-proliferative response, such as p53/TP53, LOX and CDKN1A; apoptosis, such as BBC3/PUMA, CASP1, CASP7 and CASP8; immune response, such as IL7, IL12A/B and IL15, PTGS2/COX2 and CYBB; DNA damage responses and DNA repair, such as POLQ/POLH; MHC class I expression, such as TAP1, PSMB9/LMP2, PSME1/PA28A, PSME2/PA28B and B2M and MHC class II expression, such as CIITA; metabolic enzymes, such as ACOD1/IRG1 (PubMed:15226432, PubMed:15509808, PubMed:17516545, PubMed:17942705, PubMed:18497060, PubMed:19404407, PubMed:19851330, PubMed:22367195). Represses genes involved in anti-proliferative response, such as BIRC5/survivin, CCNB1, CCNE1, CDK1, CDK2 and CDK4 and in immune response, such as FOXP3, IL4, ANXA2 and TLR4 (PubMed:18641303, PubMed:22200613). Stimulates p53/TP53-dependent transcription through enhanced recruitment of EP300 leading to increased acetylation of p53/TP53 (PubMed:15509808, PubMed:18084608). Plays an important role in immune response directly affecting NK maturation and activity, macrophage production of IL12, Th1 development and maturation of CD8+ T-cells (PubMed:11244049, PubMed:11846971, PubMed:11846974, PubMed:16932750). Also implicated in the differentiation and maturation of dendritic cells and in the suppression of regulatory T (Treg) cells development (PubMed:11244049, PubMed:11846971, PubMed:11846974, PubMed:16932750). Acts as a tumor suppressor and plays a role not only in antagonism of tumor cell growth but also in stimulating an immune response against tumor cells (PubMed:20049431).
Cell development path
A stable KO cell pool was generated using the CRISPR‑RNP approach; Sanger sequencing revealed a knockout efficiency of 97% in the cell pool.
Application
A gene knockout cell pool with high knockout efficiency (KO Cell Pool) is particularly well suited for preliminary functional analyses, the development of complex disease models, precision drug screening, and broad‑scale gene discovery studies. The KO pool can be directly applied to a variety of assays and analyses without the need for time‑consuming single‑clone selection, significantly enhancing experimental throughput.
Cell Culture Instructions
Cell Resuscitation
01. Preheat the complete culture medium in a 37°C water bath.
02. Thaw the cryovial in a 37°C water bath for 1–2 minutes.
03. Transfer the cryovials into a biosafety cabinet and wipe their surfaces with 70% ethanol.
04. Unscrew the cap of the cryovial and gently transfer the cell suspension into a sterile centrifuge tube containing 9 mL of complete culture medium.
05. Centrifuge at room temperature at 125g for 5–7 minutes, then discard the supernatant.
06. Resuspend the cell pellet in 5 mL of complete culture medium, and transfer the cell suspension to a T25 culture flask.
07. Transfer the cells to a 37°C incubator with 5% CO2 for culture.
08. Recommended passage ratio: 1/2 to 1/4; cells reach confluence in 2–3 days.
cell passage
01. When the cell confluence in the culture flask reaches 80%–90% or higher, cells can be passaged.
02. Remove the culture medium, PBS, and trypsin (0.25% Trypsin‑EDTA, Gibco 25200‑056) from the 4°C refrigerator, place them in a 37°C water bath, and once their temperature approaches 37°C, take them out. Spray the exterior of the bottles with 75% ethanol, then transfer them to a biosafety cabinet.
03. Remove the culture flask to be passaged from the incubator, spray the exterior of the flask with 75% ethanol, and place it inside a biosafety cabinet.
04. To avoid dispersing the cells, rinse the cells with PBS along the inner wall of the culture flask; discard the wash solution, then add 2 mL to a T25 flask.
05. Add the appropriate volume of trypsin (1.5 mL for a T75 flask, 0.5 mL for a T25 flask), and gently swirl the flask to ensure the enzyme evenly covers the cell‑covered bottom. Adjust the volume as needed based on the specific situation. After approximately 1–2 minutes, when most cells have detached, add an equal volume of complete culture medium to stop the digestion, then gently pipette up and down with a 5‑mL pipette until all cells are fully detached.
06. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 300 × g for 5 minutes, and discard the supernatant.
07. Transfer 5 mL of complete culture medium to resuspend the cells, adjust the seeding ratio as needed, and replenish the culture flask with complete medium—add up to 13–15 mL for a T75 flask and 5 mL for a T25 flask—then supplement with 1% double antibiotic.
08. After capping the bottle and tightening the lid, gently shake the vial to ensure uniform mixing of the cells, then place it in a 37°C incubator with 5% CO2.
cell cryopreservation
01. Prepare the cryopreservation solution and pre-cool it in advance.
02. Ensure that the cells to be cryopreserved meet the cryopreservation requirements, and verify the following conditions under a microscope: healthy appearance and morphological characteristics, being in the late logarithmic growth phase, and showing no signs of contamination or senescence.
03. Digest and centrifuge the cells (refer to the subculture protocol for specific steps).
04. Resuspend the cells in cryopreservation medium at 1 mL per tube, gently pipette up and down to ensure uniformity, then aliquot into cryovials.
05. Place the cells in a programmed cooling box and freeze them in a −80°C freezer.
06. Subsequently, the cells are transferred to a liquid nitrogen tank for long-term storage.
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