pLenti-ARMC2-sgRNA

Cat. No.: P743902

Specifications

Product Name

pLenti-ARMC2-sgRNA

Product Line

Produtos biológicos

Armazenamento e envio

Condições de armazenamento de armazenamento

Store at -80°C

Enviado em

Dry ice packs + Cold packs

Estabilidade e armazenamento

Stored at -20 ℃, valid for at least 6 months;Stored at -80 ℃, valid for at least one year.

Application Protocols

Only application details from Product Data can be provided in this section. For this item, no tested applications, recommended conditions, or protocols are specified.

• Tested applications: Not specified for this item; refer to CoA/Spec Sheet.
• Recommended dilutions/amounts: Not specified for this item; refer to CoA/Spec Sheet.
• Positive/negative controls: Not specified for this item; refer to CoA/Spec Sheet.

For general workflow guidance, please see the Reaction & Applications and Reaction Conditions sections. All uses are for research use only.

Biological Roles

Item-specific

• Target specificity details for this construct (exact spacer sequence, transcript isoform, promoter) are not specified for this item; refer to CoA/Spec Sheet.

General context (literature)

• ARMC2 (Armadillo Repeat-Containing 2): Encodes a protein with armadillo repeat motifs, a structural feature associated with protein–protein interactions. Ortholog studies and emerging literature implicate ARMC2 in ciliogenesis/flagellar function and germ cell biology, suggesting roles in motility-related structures and intracellular transport complexes. Precise cellular functions may be context- and species-dependent.
• Research utility: Targeting ARMC2 with CRISPR tools enables perturbation of pathways related to ciliary assembly, microtubule-associated processes, or reproductive cell development. Such perturbations can elucidate gene function in differentiation models, epithelial systems, or cell lines engineered for cilia studies.
• Readouts: Phenotypic assays may include cilia length/number quantification (imaging), motility measurements, transcriptomic shifts (RNA-seq), or proteomic changes in interactome analyses. At the molecular level, assessing on-target indel profiles and off-target events is critical for interpretability.

Caveats (general)

• Redundancy/compensation: Functional redundancy within armadillo repeat protein families can mask phenotypes; use multiple guides/perturbation strengths.
• Off-target effects: Employ computational design and empirical validation. Complement loss-of-function with rescue via sgRNA-resistant ARMC2 cDNA for causality.

All statements above reflect general biology/literature and are not specific performance claims for this item.

Buffer Applications

This product is a plasmid DNA construct rather than a small-molecule buffer component; classical buffer formulation guidance is not directly applicable.

General notes (literature)

• Storage/handling buffers: Nuclease-free water or TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH ~8) are commonly used to stabilize plasmid DNA during storage. EDTA chelates Mg2+ to inhibit nucleases but may inhibit Mg2+-dependent enzymes; use water for immediate enzymatic reactions when necessary.
• Working solutions: For transfection or viral packaging, plasmids are typically diluted into sterile, nuclease-free water or Opti-MEM/DMEM immediately before use.

Item-specific buffer compositions and recommended dilutions are not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

While green chemistry frameworks are most developed for small-molecule synthesis and solvent selection, several sustainable practices apply to plasmid DNA workflows.

Greener options/practices (general)

• Minimize solvent waste: Choose alcohol-efficient DNA purification methods or column kits that reduce isopropanol/ethanol use. Avoid chloroform/phenol extraction when possible; opt for spin columns or magnetic-bead systems.
• Low-waste transfection: Use high-efficiency reagents to reduce repeat experiments and consumables. Scale reactions appropriately to minimize plastic and buffer waste.
• Energy efficiency: Store working stocks at −20 °C short-term when compatible and reserve −80 °C for long-term master stocks to reduce freezer door openings and energy load.
• Non-hazardous disinfectants: For bench decontamination, use appropriately diluted bleach followed by ethanol/water rinses to limit harsh solvent residues.

Comparison (general)

• Phenol–chloroform extraction vs silica column: Columns reduce hazardous solvent handling and simplify waste, at the expense of single-use plastic. Closed-loop phenol-free systems or bead-based methods can be a balanced alternative.
• Alcohol precipitation vs bead cleanup: Bead-based cleanups often consume less alcohol and provide consistent recovery with less user exposure.

Item-specific green attributes for pLenti-ARMC2-sgRNA are not specified; refer to CoA/Spec Sheet.

Pharmaceutical Uses

This is a research-use plasmid DNA reagent and is not intended for human or veterinary use, diagnostics, or therapeutics.

General context (information only; not specific to this item)

• Plasmid DNA can serve as an intermediate in biomanufacturing research (e.g., template for viral vector development, nonclinical process development). Such activities require GMP-grade materials for clinical applications; this item’s grade is not specified and should be considered research-only.
• Regulatory considerations: Clinical or manufacturing uses would demand compliance with pharmacopeial/ICH guidelines (endotoxin, residual solvents, identity, sterility). No such compliance is claimed or specified for this item.

Item-specific pharmacopeial status, excipient roles, or formulation compatibility: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Item-specific (from Product Data)

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration, buffer, A260/280: Not specified for this item; refer to CoA/Spec Sheet.

General properties for plasmid DNA (literature)

• Physical state: Plasmid DNA is typically supplied in aqueous buffer (e.g., nuclease-free water or TE). Alternatively, it may be lyophilized.
• Solubility: Highly water soluble; insoluble in nonpolar organic solvents.
• pH stability: DNA is most stable near neutral to slightly basic pH (7.4–8.5) and is susceptible to acid hydrolysis at low pH.
• Temperature sensitivity: Repeated freeze–thaw can cause nicking; long-term storage at −80 °C is standard.
• Spectroscopy: UV λmax ~260 nm; typical A260/280 ~1.8–2.0 for clean DNA (literature).
• Viscosity: Solutions can be moderately viscous at high concentration due to polymeric nature.

Not applicable/specification note

• Melting point, boiling point, density, refractive index, logP: Not applicable to macromolecular DNA solutions or not customarily specified for plasmids.

Quality and Grades

Item-specific

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.

How to interpret grades for plasmid DNA (general guidance)

• Research-grade plasmid: Suitable for cloning, transfection, and routine functional genomics. Typical expectations (not specified for this item) include sequence verification of critical regions and low endotoxin for robust transfection.
• Endotoxin levels: Low-endotoxin plasmids (<10 EU/mg, literature) are preferred for sensitive cell types. Exact endotoxin for this item is not specified; refer to CoA.
• Sequence validation: For sgRNA constructs, identity is commonly verified by Sanger sequencing across the U6 promoter–sgRNA insert junctions. Verification status for this item is not specified; refer to CoA.
• Host strain/antibiotic marker: Usually provided on the spec sheet for propagation and selection in E. coli. Not specified for this item.
• QC assays often provided (examples): A260/280 ratio, supercoiled percentage (gel or CE), restriction digest pattern, and sterility testing. None are specified here for this item.

Practical notes

• If performing viral packaging or genome editing, request or confirm: complete plasmid map/GenBank file, antibiotic selection, backbone version, sgRNA spacer sequence, and recommended cloning sites to ensure compatibility with your workflow.

Reaction and Applications

Item-specific applications from catalog

• Manufacturer Applications: Not specified for this item; refer to CoA/Spec Sheet.

General applications for pLenti sgRNA plasmids (literature)

• CRISPR knockout (CRISPR-Cas9): pLenti sgRNA plasmids are used to express a gene-specific guide targeting ARMC2, enabling Cas9-mediated double-strand breaks and indel formation for gene disruption in mammalian cells.
• CRISPRi/CRISPRa: When used with dCas9-KRAB (repression) or dCas9-VP64/p300 (activation), the sgRNA directs transcriptional modulation of ARMC2 without cutting DNA.
• Stable delivery via lentivirus: Co-transfection with packaging plasmids in HEK293T or similar cells produces pseudotyped lentiviral particles that transduce dividing and non-dividing cells, enabling stable sgRNA expression after integration.
• Pooled screening: ARMC2-targeting guides can be part of focused libraries to interrogate ciliary/cytoskeletal functions, fertility pathways, or other processes where ARMC2 is implicated (literature).

Practical tips

• Guide validation: Confirm spacer sequence specificity with current genome builds; perform off-target prediction and select alternative guides as needed.
• Controls: Include non-targeting and safe-harbor guides (e.g., AAVS1) plus a positive control guide against an essential gene for assay benchmarking.
• Readouts: Validate editing by amplicon sequencing, TIDE/ICE analysis, or targeted NGS; confirm functional impact at RNA/protein levels.
• Packaging: Use third-generation packaging systems and perform titering (p24 ELISA or functional MOI determination) before experimental transductions.

All uses are for research use only.

Reaction Conditions

No chemical reaction conditions are applicable to this macromolecular DNA product in the traditional sense. The following guidance refers to common molecular biology workflows (literature/general):

Cloning of sgRNA spacers

• Enzymes: Type IIS restriction enzymes (e.g., BsmBI/BbsI) and T4 ligase, or Gibson/HiFi assembly.
• Conditions: 1× manufacturer buffer; digest 15–60 min at 37 °C; ligation 10–30 min at room temperature or 16 °C overnight. Transform into high-efficiency competent E. coli.

Lentiviral packaging (BSL-2, institutional approvals required)

• Cells: HEK293T or derivatives at ~70–90% confluence.
• Transfection: Use PEI, lipids, or calcium phosphate per vendor instructions; typical DNA ratios follow the packaging system (e.g., 4:3:1 transfer:packaging:envelope by mass; literature). Replace media after 6–18 h.
• Harvest: Collect supernatant at 48–72 h; clarify by low-speed spin and 0.45 µm filtration. Optional concentration by PEG or ultracentrifugation.

Transduction and selection

• MOI: Optimize empirically; spinoculation can enhance efficiency. Include polybrene (e.g., 4–8 µg/mL) if compatible with cells.
• Selection: Begin 48–72 h post-transduction if a selectable marker is present; duration depends on kill curve.

Editing validation

• Extract genomic DNA 3–7 days post-transduction; PCR amplify target; quantify editing by TIDE/ICE or targeted NGS.

Note: Parameters above are general literature guidance and not specifications for this item.

Safety and Handling

Item-specific hazard information

• GHS classification, signal word, hazard/pictograms, H-statements: Not specified for this item; refer to the SDS.

General laboratory safety for plasmid DNA and lentiviral vectors (literature/typical)

• Biosafety: Purified plasmid DNA itself is non-infectious. However, when used to produce lentiviral particles (via co-transfection with packaging plasmids), all work should follow institutional BSL-2/BSL-2+ practices and approvals. Decontaminate benches and tools with 10% bleach followed by ethanol.
• PPE: Laboratory coat, nitrile gloves, and safety glasses. Change gloves frequently to avoid nuclease contamination.
• Handling: Use nuclease-free tubes and tips. Avoid repeated freeze–thaw; aliquot upon first thaw to maintain integrity. Keep on ice during set-up of molecular biology workflows.
• Storage incompatibilities: Avoid strong acids/bases and nucleases (RNases/DNases). Keep away from microbial contamination; work in a clean area.
• First-aid (general): In case of accidental contact, rinse skin/eyes with water. If inhaled as aerosol from downstream viral work, follow institutional exposure protocols. Seek medical attention as appropriate.
• Waste: Non-recombinant plasmid waste is typically disposed as biohazardous solid/liquid per local regulations. Lentiviral production waste must be disinfected (e.g., bleach) prior to disposal per BSL guidelines.

Authoritative guidance

• Always consult the product SDS and your institutional biosafety committee policies for definitive safety requirements.

Solvent Selection

This product is a plasmid DNA reagent rather than a small-molecule solute; conventional organic solvent selection is not typically applicable.

General guidance for handling DNA solutions (literature)

• Solvent system: Use nuclease-free water or TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0). TE offers enhanced long-term stability by buffering pH and chelating divalent cations that can catalyze hydrolysis.
• Avoid: Organic solvents (e.g., alcohols, chlorinated solvents) for dissolution. Short alcohol exposures are used only in precipitation/wash steps during purification, not for storage.
• Compatibility: DNA solutions are compatible with common aqueous biological buffers (PBS, HBSS) for certain applications, but high-salt buffers may affect transfection efficiency.
• Filtration: If sterile filtration is necessary, use low protein/DNA-binding membranes (e.g., PVDF or PES, 0.22 µm) and minimize shear.

When to choose water vs TE

• Water: For immediate use in enzymatic reactions sensitive to EDTA (e.g., certain ligations or transcriptions).
• TE buffer: For long-term storage to maximize stability and nuclease resistance.

Item-specific buffer composition for pLenti-ARMC2-sgRNA is not specified; refer to CoA/Spec Sheet.

Storage and Reconstitution

Item-specific (from Product Data)

• Storage conditions: Store at −80 °C.
• Shipping: Shipped on dry ice packs + cold packs.

Additional item-specific details

• Appearance, buffer, concentration, and whether the product is lyophilized or in solution: Not specified for this item; refer to CoA/Spec Sheet.

General guidance for plasmid DNA (literature; not item-specific specifications)

• Aliquoting: Upon first receipt, thaw on ice, mix gently (do not vortex), briefly spin, and aliquot into nuclease-free tubes to avoid repeated freeze–thaw cycles.
• Short-term storage: Working aliquots can be kept at −20 °C for weeks to months; minimize freeze–thaw.
• Long-term storage: Maintain master stocks at −80 °C as indicated. TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH ~8) supports stability.
• Reconstitution (if lyophilized): Reconstitute with nuclease-free water or TE to the desired concentration; allow to hydrate on ice for 10–15 min; mix gently by pipetting.
• Handling: Use low-binding tips/tubes; keep on ice during setup. Verify concentration by A260 (dsDNA: 50 µg/mL per A260 = 1, literature) and assess integrity by agarose gel if needed.

Research use note

• For research use only.

Structure and Identity

Item-specific (from Product Data)

• Product name: pLenti-ARMC2-sgRNA (SKU: P743902)
• CAS: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey / SMILES: Not applicable to plasmid DNA constructs.

General description (literature/typical for this class)

• Construct type: Circular double-stranded DNA plasmid on a lentiviral backbone designed to express a single-guide RNA (sgRNA) targeting the ARMC2 gene.
• Typical structural features in pLenti sgRNA vectors: U6 or H1 Pol III promoter driving the sgRNA, sgRNA scaffold (tracrRNA-derived), optional selection marker (e.g., puromycin resistance), lentiviral LTRs and packaging signal (Ψ) for virus production in packaging cells, and bacterial origin of replication plus antibiotic resistance for propagation in E. coli.
• 2D/functional layout (textual): [5' LTR] – [Ψ packaging signal] – [U6 promoter] – [sgRNA spacer targeting ARMC2] – [sgRNA scaffold] – [selection marker cassette (varies)] – [bacterial ori + antibiotic resistance] – [3' LTR]. Exact map and elements vary by specific plasmid version and are not specified for this item; refer to CoA/Spec Sheet.
• Stereochemistry: Not applicable (nucleic acid polymer).

Synthetic Utility

Although not a small-molecule building block, a pLenti sgRNA plasmid is a versatile synthetic biology tool enabling programmable genome manipulation.

General utilities (literature)

• Modular guide exchange: Most pLenti sgRNA vectors accept short oligos (~20 nt spacer) by Golden Gate cloning (e.g., BsmBI/BbsI). This allows rapid retargeting to different loci, including ARMC2 isoforms or regulatory regions.
• Cassette engineering: Backbones support insertion of alternative promoters (H1 vs U6), dual-guide arrays, or expression cassettes for fluorescent markers/selection, enabling streamlined selection and tracking.
• Combinatorial perturbations: Co-expression with Cas9 or dCas9 fusions facilitates knockout, repression, or activation without altering the backbone chemistry; multiplexing supports pathway mapping.
• Retrosynthetic workflow (conceptual): Desired phenotype -> identify ARMC2 target site(s) -> design on-target, low off-target spacers -> clone into pLenti backbone -> validate by Sanger -> package into lentivirus -> transduce -> select -> assay -> rescue/control experiments.

Practical considerations

• Sequence constraints: U6 promoters prefer a 5'-G; adapt designs or append G when necessary (not always required depending on promoter architecture).
• Backbone compatibility: Confirm antibiotic selection and origin of replication for strain choice and copy number. Verify presence/absence of cPPT/CTS or WPRE if expression tuning is relevant.

Item-specific backbone map and cloning sites are not specified; refer to CoA/Spec Sheet.

Target Specificity

Only information derived from Product Data can be listed here.

• Antigen/gene target: Not specified for this item; refer to CoA/Spec Sheet. (Product name suggests ARMC2 as the intended gene target, but the exact spacer sequence and isoform are not provided.)
• Species reactivity/target genome build: Not specified for this item; refer to CoA/Spec Sheet.
• Clone/construct identifiers, promoter, selection marker: Not specified for this item; refer to CoA/Spec Sheet.
• Validations or tested applications: Not specified for this item; refer to CoA/Spec Sheet.