SPPL3 Knockout HEK293 Cell Line
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Cat.No.:
EDC07590
Species:
Human
Cell Name:
HEK293
Gene:
SPPL3
Gene ID:
121665
Size:
1×10⁶cells
SPPL3 Knockout Cell Line (HEK293) is an exclusive upgraded CRISPR/Cas9 system-mediated gene knockout cell, with the advantages of Optimized Strategy Design, Efficient Cell Transfection, High-Performance Cas9 Protein and Hassle-Free Cell Selection.
| Cat.No. | EDC07590 |
|---|---|
| Product Name | SPPL3 Knockout Cell Line (HEK293) |
| Cell Line | HEK293 |
| Cellosaurus ID | CVCL_0045 |
| Cell Line Synonyms | Hek293, HEK-293, HEK/293, (HEK)293, HEK 293, HEK,293, 293, 293 HEK, 293 Ad5, Graham 293, Graham-293, Human Embryonic Kidney 293 |
| Gene | SPPL3 |
| NCBI Gene ID | |
| Gene Synonyms | IMP2|MDHV1887|PRO4332|PSH1|PSL4 |
| Summary |
Enables aspartic endopeptidase activity, intramembrane cleaving and protein homodimerization activity. Involved in T cell receptor signaling pathway; membrane protein proteolysis; and positive regulation of calcineurin-NFAT signaling cascade. Located in Golgi-associated vesicle membrane; endoplasmic reticulum; and plasma membrane. [provided by Alliance of Genome Resources, Jul 2025]
|
| Associated Diseases | Non-tumor |
| Morphology | Adherent |
| Passage Ratio | 1/5，2days |
| Complete Culture Medium | DMEM + 10% FBS |
| Freezing Medium | 95% Complete culture medium+ 5% DMSO |
| QC | Indels validated by Sanger sequencing; sterility confirmed via microbial testing. |
* For research use only. Not intended for use in humans or animals, including clinical, therapeutic, or diagnostic purposes.
| Loci | STR Info (Sample Cell) Sample Cell Line: HEK293 | STR Info (Cell bank) Cell Line: HEK293 | ||
| Allele1 | Allele2 | Allele1 | Allele2 | |
| Amelogenin | X | X | ||
| CSF1P0 | 12 | 11 | 12 | |
| D2S1338 | 19 | 19 | ||
| D3S1358 | 15 | 17 | 15 | 17 |
| D5S818 | 8 | 8 | 9 | |
| D7S820 | 11 | 12 | 11 | 12 |
| D8S1179 | 12 | 14 | 12 | 14 |
| D13S317 | 12 | 14 | 12 | 14 |
| D16S539 | 9 | 13 | 9 | 13 |
| D18S51 | 17 | 18 | 17 | 18 |
| D19S433 | 15 | 18 | 15 | 18 |
| D21S11 | 28 | 30.2 | 28 | 30.2 |
| FGA | 23 | 23 | ||
| Penta D | 9 | 10 | 9 | 10 |
| Penta E | 7 | 15 | 7 | 15 |
| TH01 | 7 | 9.3 | 7 | 9.3 |
| TPOX | 11 | 11 | ||
| vWA | 16 | 19 | 16 | 19 |
| D6S1043 | 11 | 11 | ||
| D12S391 | 19 | 21 | 11 | 15 |
| D2S441 | 11 | 15 | 11 | 15 |
* STR authentication data of this cell line matches with that of cell lines sourced from ATCC, DSMZ, JCRB, and RIKEN databases.
Conclusion: The STR identification of this cell is correct.
Conclusion: The STR identification of this cell is correct.
FAQ
Which is better for studying SPPL3 function, SPPL3 Knockout HEK293 Cell Line or SPPL3 overexpression HEK293 Cell Line?
The choice depends on whether you are studying SPPL3's role as a non-canonical signal peptide peptidase or its emerging functions in glycoprotein homeostasis and ectodomain shedding. The Knockout line is appropriate for asking whether SPPL3 is required for proteolytic processing of glycosyltransferases (B4GALT1, ST6GAL1, others) — a function that distinguishes SPPL3 from related SPPL proteases. Overexpression is useful for testing whether elevated SPPL3 drives enhanced ectodomain shedding of these substrates.
For SPPL family research, the EDITGENE SPPL3 Knockout in HEK293 is the standard mechanistic platform — HEK293 has been extensively used for SPPL substrate characterization and structural studies. Rescue with wild-type or catalytically-dead SPPL3 is the standard specificity control. The role of SPPL3 in modifying cellular glycoprofile makes the knockout particularly valuable for glycobiology research.
What are the application scenarios for this model?
Primary applications:
• Glycoprotein ectodomain shedding: Western blot analysis of glycosyltransferase substrates (B4GALT1, ST6GAL1, MGAT5) to assess SPPL3-dependent proteolysis.
• Cellular glycoprofile analysis: lectin staining (RCA-I, MAL-II, PHA-L) or glycomic analysis to detect SPPL3 loss-induced glycosylation changes.
• Substrate identification: secretome proteomics to identify novel SPPL3 substrates beyond the known glycosyltransferase set.
• In vitro protease activity: SPPL3 catalytic activity assessment using recombinant or immunoprecipitated enzyme on candidate substrates.
EDITGENE recommends this model for researchers investigating signal peptide peptidase-like proteases, glycoprotein homeostasis, and SPPL substrate biology.
Is this SPPL3 Knockout HEK293 Cell Line compatible with overexpression rescue experiments?
Yes. SPPL3 rescue experiments require attention to membrane topology and protease activity:
• Construct design: use a codon-modified SPPL3 sequence with a cytoplasmic C-terminal tag (FLAG, HA). SPPL3 is a multi-pass membrane intramembrane aspartyl protease — the catalytic YD and GxGD motifs in the membrane domain must be preserved.
• Catalytically-dead rescue: mutations in the catalytic aspartates (D205A or D271A in conserved YD or GxGD motifs) abolish protease activity and serve as the standard specificity control.
• Substrate-specific rescue: rescue should restore proteolytic processing of glycosyltransferase substrates (assessed by Western blot or glycoprofile analysis).
• Localization validation: confirm Golgi localization of exogenous SPPL3 by immunofluorescence co-staining with GM130 before functional assays.
HEK293 transduces efficiently with lentivirus and supports SPPL3 expression at levels enabling biochemical characterization.
* Research Use Disclaimer: Content is generated from publicly available research data, bioinformatic resources, and computational analyses for research reference only.
Related Publications
Signal peptide peptidase- and SPP-like 3-dependent shedding of α1,6-fucosyltransferase differentially affects core fucosylation.
IF=3.9
The Journal of biological chemistry
Alpha1,6-fucosyltransferase (FUT8) biosynthesizes core fucose on N-glycans, which plays essential roles in various biological processes, including immunity and development. Although FUT8 is a Golgi-resident type II membrane protein, it is also secreted by an unknown mechanism. Here, we demonstrate that signal peptide peptidase (SPP) and signal peptide peptidase-like 3 (SPPL3), members of an intramembrane protease family, both cleave FUT8 for secretion. Knockout of SPP or SPPL3 in cells partially impaired FUT8 secretion, and double KO led to more drastic impairment in secretion, indicating that SPP and SPPL3 independently cleave FUT8. Sequencing analysis revealed that the N terminus of FUT8 in the media was mapped in the stem region, which is far from the expected cleavage site for SPP/SPPL3, suggesting that FUT8 undergoes two-step proteolytic processing, initially by SPP/SPPL3 and subsequently by another protease. Moreover, glycoproteomics suggested that the substrate glycoprotein preference of FUT8 was altered by knocking out SPP or SPPL3, highlighting the importance of FUT8 shedding in core fucosylation.
This KO model may be useful for:
- Investigating the role of SPPL3 in regulated intramembrane proteolysis (RIP) of glycosylation enzymes
- Studying the impact of SPPL3-dependent shedding of α1,6-fucosyltransferase on core fucosylation
- Elucidating the molecular mechanisms linking SPPL3 activity to N-glycan maturation and modification
- Exploring the functional consequences of altered core fucosylation in cell signaling and adhesion
- Providing a cellular platform for screening modulators of SPPL3-mediated substrate cleavage
Required Accessories
Cell Culture Optimization Series
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