{"product_id":"aifm1-crispr-knockout-l929-stable-cell-line-negative-clone-t2-15-3-bhc10900953","title":"AIFM1 CRISPR Knockout L929 Stable Cell Line - Clone T2-15-3","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eThis product is a stable CRISPR knockout cell line in which \u003cem\u003eAifm1\u003c\/em\u003e (Apoptosis Inducing Factor Mitochondria Associated 1) has been disrupted in the L929 mouse fibroblast background. Generated using CRISPR-Cas9 technology, this line provides an isogenic model for studying the role of AIFM1 in apoptosis, mitochondrial function, and redox biology. Cells are supplied frozen (1×10⁶ cells \/ 1.0 ml), classified as BSL-II, and grown as adherent, fibroblast-like monolayers.\u003c\/p\u003e\n\n\u003ch2\u003eCRISPR Knockout Design\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eTarget gene:\u003c\/strong\u003e \u003cem\u003eAifm1\u003c\/em\u003e (Apoptosis Inducing Factor Mitochondria Associated 1; mouse gene symbol)\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eKnockout strategy:\u003c\/strong\u003e CRISPR-Cas9–mediated disruption of the \u003cem\u003eAifm1\u003c\/em\u003e locus with stable integration confirmed by puromycin selection\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSelection marker:\u003c\/strong\u003e Puromycin resistance; 7.0 µg\/ml puromycin used for selection (add only after first passage post-thaw)\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eParental cell line:\u003c\/strong\u003e L929 — mouse fibroblast cell line derived from normal subcutaneous areolar and adipose connective tissue (Male, C3H\/An, 100 days); adherent, fibroblast-like morphology\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eClone ID:\u003c\/strong\u003e T2-15-3 (negative clone designation)\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eGene Background\u003c\/h2\u003e\n\u003cp\u003eAIFM1 (Apoptosis Inducing Factor Mitochondria Associated 1), also known as AIF, is a mitochondrial flavoprotein encoded by a nuclear gene and imported into the mitochondrial intermembrane space. Under normal physiological conditions, AIFM1 is involved in maintaining mitochondrial morphology and supporting oxidative phosphorylation through NADH oxidase activity. Upon receipt of apoptotic signals, AIFM1 is cleaved and released from the inner mitochondrial membrane, translocates to the nucleus, and triggers large-scale DNA fragmentation and chromatin condensation through a caspase-independent mechanism. Mutations and dysregulation of AIFM1 have been associated with mitochondrial encephalomyopathy and Cowchock syndrome, and the protein has been studied in the context of neurodegeneration, cancer cell death, and ischemia-reperfusion injury. The mouse \u003cem\u003eAifm1\u003c\/em\u003e gene is located on the X chromosome.\u003c\/p\u003e\n\n\u003ch2\u003eCell Culture Specifications\u003c\/h2\u003e\n\u003cdl\u003e\n  \u003cdt\u003eRecommended vessels\u003c\/dt\u003e\n\u003cdd\u003ePriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422)\u003c\/dd\u003e\n  \u003cdt\u003eCulture medium\u003c\/dt\u003e\n\u003cdd\u003eRPMI 1640 Medium (TM503) + 10% FBS (do not heat-inactivate) + 1% Penicillin\/Streptomycin Solution (G255)\u003c\/dd\u003e\n  \u003cdt\u003eCulture conditions\u003c\/dt\u003e\n\u003cdd\u003e37.0°C, 5% CO₂\u003c\/dd\u003e\n  \u003cdt\u003eSelection\u003c\/dt\u003e\n\u003cdd\u003e7.0 µg\/ml Puromycin (G264); add only after first passage post-thaw\u003c\/dd\u003e\n  \u003cdt\u003eGrowth properties\u003c\/dt\u003e\n\u003cdd\u003eAdherent, fibroblast-like\u003c\/dd\u003e\n  \u003cdt\u003eOrganism\u003c\/dt\u003e\n\u003cdd\u003eMouse (\u003cem\u003eM. musculus\u003c\/em\u003e)\u003c\/dd\u003e\n  \u003cdt\u003eTissue origin\u003c\/dt\u003e\n\u003cdd\u003eConnective Tissue (subcutaneous areolar and adipose tissue)\u003c\/dd\u003e\n  \u003cdt\u003eBiosafety level\u003c\/dt\u003e\n\u003cdd\u003eBSL-II\u003c\/dd\u003e\n  \u003cdt\u003eFormat\u003c\/dt\u003e\n\u003cdd\u003eFrozen; 1×10⁶ cells \/ 1.0 ml\u003c\/dd\u003e\n\u003c\/dl\u003e\n\u003cp\u003eKnockout is confirmed by the presence of frameshift-inducing INDELs, verified by Sanger Sequencing.\u003c\/p\u003e\n\n\u003ch2\u003eResearch Applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eApoptosis pathway analysis:\u003c\/strong\u003e Investigate caspase-independent cell death mechanisms by comparing Aifm1 KO L929 cells to the isogenic parental line under apoptotic stimuli (e.g., H₂O₂, staurosporine, serum withdrawal).\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eMitochondrial function studies:\u003c\/strong\u003e Profile mitochondrial membrane potential (ΔΨm), ATP production, and oxygen consumption rate (OCR) in the absence of functional AIFM1 using standard respirometry assays.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eDrug sensitivity screening:\u003c\/strong\u003e Screen compounds targeting the intrinsic apoptotic pathway in the Aifm1 KO background; use the L929 parental line as the isogenic control.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eRedox biology:\u003c\/strong\u003e Quantify intracellular ROS levels and oxidative stress markers (e.g., lipid peroxidation, glutathione) in AIFM1-deficient cells.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eRescue experiments:\u003c\/strong\u003e Re-express wild-type or mutant AIFM1 constructs in the KO background to investigate structure-function relationships.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eImportant Notes\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSelection drug timing:\u003c\/strong\u003e Add puromycin (7.0 µg\/ml) to culture medium only after the first passage post-thaw to allow cells to recover from freeze-thaw conditions.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eDonor\/source:\u003c\/strong\u003e Parental L929 cells are derived from normal subcutaneous areolar and adipose connective tissue of a male C3H\/An mouse (approximately 100 days of age).\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eViability warranty:\u003c\/strong\u003e Cell viability upon thaw is warranted for 30 days following shipment when handled according to abm's cell handling guidelines.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eIntended use:\u003c\/strong\u003e For laboratory research use only. Not intended for diagnostic, therapeutic, or clinical applications.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003c!-- Sources (internal):\n- AIFM1 gene entry | UniProt (Q9Z0X1 mouse) | https:\/\/www.uniprot.org\/uniprot\/Q9Z0X1\n- Aifm1 gene record | NCBI Gene ID: 26926 | https:\/\/www.ncbi.nlm.nih.gov\/gene\/26926\n- Susin SA et al. (1999) Molecular characterization of mitochondrial apoptosis-inducing factor. Nature | https:\/\/doi.org\/10.1038\/46522\n- Sevrioukova IF (2011) Apoptosis-inducing factor: structure, function, and redox regulation. Antioxid Redox Signal | https:\/\/doi.org\/10.1089\/ars.2010.3445\n- Joza N et al. (2001) Essential role of the mitochondrial apoptosis-inducing factor in programmed cell death. Nature | https:\/\/doi.org\/10.1038\/35071141\n--\u003e","brand":"Applied Biological Materials (abm) Inc.","offers":[{"title":"1x10\u003csup\u003e6\u003c\/sup\u003e cells \/ 1.0 ml","offer_id":53180707635565,"sku":"T9674","price":0.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/t9674.png?v=1774960835","url":"https:\/\/www.ebiohippo.com\/products\/aifm1-crispr-knockout-l929-stable-cell-line-negative-clone-t2-15-3-bhc10900953","provider":"BioHippo","version":"1.0","type":"link"}