pLenti-VGF-sgRNA

Cat. No.: P751726

Spezifikationen

Product Name

pLenti-VGF-sgRNA

Product Line

Biologische Produkte

Lagerung und Versand

Storage

Store at -80°C

Verschickt in

Dry ice packs + Cold packs

Stabilität und Lagerung

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

Application Protocols

No tested applications, recommended dilutions, or validated protocols are provided in the Product Data for this item.

Please refer to the CoA/Spec Sheet for any item-specific instructions. Users may follow institutionally approved, general CRISPR lentiviral workflows for packaging, transduction, and selection, but these are not validated for this specific product in the provided documentation.

Biological Roles

Target gene context: VGF (nonacronymic) encodes a secreted precursor protein that is processed into multiple bioactive neuropeptides. It is widely studied in neuronal and neuroendocrine systems for roles in synaptic function, energy balance, and neurotrophic signaling (literature). This reagent’s sgRNA is intended to direct Cas nuclease activity to the VGF locus for perturbation studies.

Research applications enabled (general knowledge)

• Gene knockout/knockdown: Disruption of VGF coding sequence or regulatory regions to assess effects on peptide processing and cellular phenotypes.
• Transcriptional modulation: With dCas9 effectors, sgRNAs targeting promoter/enhancer regions can downregulate (CRISPRi) or upregulate (CRISPRa) VGF expression to probe pathway dependencies.
• Pathway analysis: Examine downstream effects on neuropeptide secretion, cAMP signaling, synaptic plasticity markers, or metabolic readouts linked to VGF-regulated biology.

Experimental considerations

• Cellular context: Choose relevant neuronal or endocrine cell lines/primary cells that express VGF for meaningful phenotypic readouts.
• Redundancy and specificity: Utilize multiple independent sgRNAs against VGF to validate on-target effects and rule out idiosyncratic off-targets.

Note: No species, isoform, or exact sgRNA sequence details are provided in the Product Data; consult CoA/Spec Sheet for target sequence and alignment information if available.

Buffer Applications

Not typically applicable as a stand-alone buffer system. This product is a lentiviral sgRNA vector.

Practical notes

• Handling buffer: If supplied in aqueous buffer, maintain nuclease-free conditions; avoid divalent cation contamination that may promote nuclease activity. Exact buffer composition is not specified for this item; refer to CoA/Spec Sheet.
• Downstream assays: When preparing lysates or nucleic acids from edited cells, select buffers compatible with your detection method (e.g., PCR lysis buffers, RIPA for protein), but these are general molecular biology considerations rather than attributes of this product.

Green Alternatives

Green chemistry substitution is not directly applicable to nucleic-acid-based reagents. However, sustainability can be improved by optimizing upstream and downstream processes.

Greener practice considerations (general knowledge)

• Minimize single-use plastics: Employ reusable, sterilizable labware where biosafety allows; consolidate aliquots to reduce waste while avoiding repeated freeze–thaw.
• Energy efficiency: Plan batch experiments to reduce frequent ultra-low temperature door openings; use insulated secondary containers when transporting on dry ice.
• Safer reagents: Prefer non-halogenated disinfectants where appropriate; avoid unnecessary solvent use in plasmid prep cleanups.
• Waste reduction: Titrate viral doses (MOI) carefully to avoid excess production; scale packaging runs to actual needs.

Tradeoffs

• Biosafety and sterility requirements limit some reuse options. Always prioritize containment and compliance over waste reduction.

These are general operational suggestions; no item-specific greener alternative exists for the pLenti-VGF-sgRNA reagent.

Pharmaceutical Uses

This reagent is provided strictly for research use only. It is not intended for diagnostic, therapeutic, or clinical manufacturing purposes.

General context (non-clinical)

• Tool compound/reagent role: Employed in discovery-stage research to interrogate gene function, validate targets, and build edited cell models. Any transition to clinical-grade vectors requires comprehensive redesign, GMP production, extensive characterization, and regulatory oversight.

Item-specific note

• No pharmacopeial status, excipient role, or clinical formulation information is provided or implied for this product.

Physical Properties

As a DNA-based biological reagent, conventional small-molecule physical property fields are generally not applicable. Where relevant values are absent in the Product Data, they are noted below.

Item-specific properties

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration, buffer, or titer: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• pH/osmolality: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general context for plasmid DNA reagents (not item-specific)

• Physical form: Typically supplied as aqueous DNA solution (e.g., TE or nuclease-free water) or as viral supernatant/aliquots for lentiviral formats.
• Stability: DNA solutions are stable when stored at −20 to −80°C; repeated freeze–thaw can degrade nucleic acids.
• Solubility: Plasmid DNA is water-soluble; insoluble in most organic solvents.
• UV properties: DNA shows a characteristic A260 peak; A260/A280 ratio ~1.8–2.0 indicates high purity (literature).

Note: Definitive specifications for this item (exact backbone, insert length, solvent/buffer, and concentration) are not provided in the Product Data and should be confirmed on the CoA/Spec Sheet.

Quality and Grades

Item-specific quality grade and purity metrics are not provided in the Product Data and should be confirmed on the CoA/Spec Sheet.

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.
• Intended use: For research use only (as stated in Product Data).

General notes on quality considerations for CRISPR lentiviral sgRNA reagents (general knowledge)

• Plasmid DNA quality: For cloning, Sanger verification, and transfection, high-purity, endotoxin-reduced DNA is typically preferred to improve cell viability and transfection efficiency. For viral production, endotoxin levels and host genomic DNA contamination should be minimized.
• Sequence verification: sgRNA spacer sequence and backbone junctions are usually verified by Sanger sequencing; full plasmid NGS may be used for critical applications.
• Functional QC: Titer (for viral stocks), MOI determination, and edit efficiency (e.g., T7E1, ICE/TIDE, or NGS) are common functional metrics, but no item-specific values are provided here.

Implications for users

• Without explicit grade specs, users should verify suitability via in-house QC (restriction digest, sequencing, bacterial propagation performance, or small-scale packaging test). Consult the product CoA/Spec Sheet for any provided sequence maps or QC data.

Reaction and Applications

This reagent is designed for CRISPR gene-editing workflows targeting the VGF locus using an sgRNA expressed from a lentiviral (pLenti) backbone.

Typical applications (general knowledge for this class)

• Stable delivery of sgRNA: Lentiviral transduction enables persistent sgRNA expression in dividing and non-dividing cells, facilitating knockout or modulation when co-expressed with a compatible Cas nuclease (e.g., SpCas9).
• Functional genomics: Generation of VGF loss-of-function lines to interrogate gene function in neuronal and endocrine models; pooled screens if incorporated into libraries (this item appears as a single sgRNA, not a library; confirm via CoA).
• Complementary modalities: Use with dCas9-KRAB for CRISPRi at VGF regulatory regions or dCas9-VP64 for CRISPRa (depending on experimental design and available constructs).

Practical considerations

• Packaging: Co-transfect with helper plasmids (e.g., gag/pol, Rev, VSV-G) into HEK293T packaging cells to produce pseudotyped lentivirus. Optimize DNA ratios and transfection method (PEI, lipids) for titer. Specific helper plasmids are not defined in this Product Data.
• Target design: Verify the sgRNA protospacer sequence, PAM compatibility, and predicted on/off-target profile; consider multiple sgRNAs to ensure robust knockout.
• Validation: Confirm VGF editing by PCR amplicon sequencing, ICE/TIDE analysis, or targeted NGS; corroborate with protein or transcript assays.

Note: No item-specific sequence, promoter, or marker details are provided; consult the CoA/Spec Sheet.

Reaction Conditions

No chemical reaction conditions apply in the traditional sense. For CRISPR workflows, the following are general, literature-based operational parameters; they are not item-specific specifications.

General operational guidance (literature/general)

• Lentiviral production: Transiently transfect HEK293T packaging cells at 40–80% confluency with transfer vector (this item), gag/pol, Rev, and VSV-G plasmids. Use serum-free complexation (e.g., PEI or lipid) and replace media after 6–12 h. Harvest supernatant at 48–72 h. Filter through 0.45 µm; concentrate if needed.
• Transduction: Apply virus to target cells with polybrene (e.g., 4–8 µg/mL) if compatible; spinoculation can enhance delivery. Select with the appropriate antibiotic or FACS if a fluorescent marker is present (marker not specified for this item).
• Editing window: Allow 48–96 h post-transduction for indel formation with active Cas9, then assay by PCR/NGS. For stable lines, expand under selection before readout.
• Controls: Include non-targeting sgRNA controls and multiple independent VGF sgRNAs to confirm specificity.

Note: Exact sgRNA sequence, promoters, resistance markers, and any titer/concentration for this product are not specified; consult the CoA/Spec Sheet. Follow institutional biosafety protocols for all steps.

Safety and Handling

Hazard information provided for this item is limited; always consult the product SDS and local biosafety officer for authoritative guidance.

Item-specific hazard data

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

General biosafety guidance for lentiviral CRISPR vectors (general knowledge)

• Biosafety level: Work with lentiviral packaging, pseudotyped particles, or transduced cells should follow BSL-2 practices at minimum, with engineering controls per institutional policy.
• PPE: Laboratory coat, gloves, and eye protection; use a certified biosafety cabinet for manipulations that may generate aerosols.
• Incompatibilities: Avoid nucleases (RNase/DNase) contamination; maintain nuclease-free technique.
• Decontamination: Use freshly prepared 10% bleach for surfaces and waste, followed by appropriate neutralization; dispose of biohazardous waste according to regulations.
• First aid overview: On exposure, follow institutional exposure control plan; flush affected area with water, seek medical evaluation as required.
• Transport and storage: Ship on dry ice/cold packs as provided; maintain −80°C storage to preserve integrity.

Special risks and notes

• Replication-incompetent vectors still require containment. Do not aerosolize. Ensure proper training for viral packaging/transduction. For DNA-only plasmid handling (no viral packaging), standard molecular biology precautions apply.

Always defer to the SDS/IBCs for definitive safety requirements.

Solvent Selection

Not typically applicable. This is a nucleic acid/viral vector reagent rather than a small-molecule solute requiring organic solvent selection.

Practical notes

• Aqueous handling: Use nuclease-free water or appropriate low-salt buffer (e.g., TE) as recommended in the CoA/Spec Sheet (not specified for this item).
• Avoid organic solvents: DNA and viral particles are generally incompatible with most organic solvents and dehydrating conditions.

For experimental planning, focus on the sections Safety & Handling, Reaction & Applications, and Storage & Reconstitution.

Storage and Reconstitution

Item-specific conditions (from Product Data)

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

General handling guidance for nucleic-acid/viral vector reagents (not item-specific)

• Thawing: Thaw on ice. Mix gently by inversion; avoid vigorous vortexing to prevent shearing or loss of viral activity.
• Aliquoting: Upon first thaw, aliquot into single-use volumes to minimize freeze–thaw cycles, which can degrade DNA or reduce viral titer.
• Temperature control: Keep on ice during setup; return unused aliquots promptly to −80°C.
• Nuclease control: Use nuclease-free tubes and tips; avoid repeated access that increases contamination risk.

Reconstitution

• If supplied lyophilized or as a dry pellet (not specified), reconstitute with nuclease-free water or the buffer recommended on the CoA/Spec Sheet. Gently pipette to dissolve; allow complete resuspension on ice.

Documentation

• Because buffer composition, concentration, and format are not specified for this item, consult the CoA/Spec Sheet for any additional storage notes (e.g., presence of cryoprotectants) and for stability upon short-term storage at −20°C or 4°C, if applicable.

Structure and Identity

This product is a genetic reagent (lentiviral CRISPR vector harboring an sgRNA) rather than a small-molecule chemical. Conventional small-molecule identifiers are not applicable.

• Item name: pLenti-VGF-sgRNA (SKU: P751726)
• Modality: Lentiviral CRISPR/Cas guide RNA vector (plasmid/viral backbone; sgRNA targeting the VGF gene)
• Category context: Listed under antibodies in catalog navigation, but this item is a nucleic-acid-based gene-editing reagent.

Item-specific identifiers (from Product Data)

• CAS: Not specified for this item; refer to CoA/Spec Sheet.
• CID: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not applicable to nucleic acid constructs; not provided.
• SMILES: Not applicable to nucleic acid constructs; not provided.
• Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.

Structural description (general knowledge for this class)

• Typical features: A lentiviral backbone (e.g., pLenti-derived) containing a U6 or H1 promoter-driven single guide RNA cassette with a 20-nt protospacer targeting VGF upstream of a scaffold. Packaging elements (LTRs, psi, RRE, cPPT) and selectable markers/promoters may be present depending on the exact backbone design (not specified here).
• 2D “structure” description: Double-stranded plasmid DNA circular vector; the functional moiety is the short sgRNA sequence expressed in cells to guide Cas nuclease to the VGF locus.

Synthetic Utility

Classically defined organic synthesis utility does not apply. However, in a genetic engineering context, the construct functions as a programmable nuclease guide to orchestrate sequence-specific genome editing at the VGF locus.

Utility in molecular biology (general knowledge)

• Modular cloning: The sgRNA cassette can be swapped or re-cloned using standard restriction/Golden Gate/Gibson methods to target alternative VGF sites or introduce barcodes (subject to the actual backbone, not specified here).
• Vector engineering: Promoters, selection markers, and reporter cassettes can be tailored to cell type and readout requirements.
• Combinatorial editing: Co-transduction with additional sgRNAs enables multiplexed perturbations to dissect pathway interactions.

Note: Specific restriction map, antibiotic resistance marker, and promoter details are not provided in the Product Data; consult the CoA/Spec Sheet for vector map if available.

Target Specificity

From Product Data naming: the sgRNA targets the VGF gene. No further target details are provided.

Item-specific details

• Target gene: VGF (as indicated by product name pLenti-VGF-sgRNA).
• Species, transcript/isoform, exact protospacer, and PAM: Not specified for this item; refer to CoA/Spec Sheet.

General considerations (not item-validated)

• On-target validation typically includes amplicon sequencing and phenotype rescue experiments.
• Off-target risk should be assessed using in silico prediction tools and empirically with unbiased methods if critical.

Only the gene target name is inferable from the Product Data provided.

Häufig gestellte Fragen

How should this product be stored?

Store at ?80 °C. Ultra-low temperature storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships frozen with dry ice and cold packs. Unpack on arrival and transfer it to the storage condition stated above; handle dry ice with insulated gloves in a ventilated area.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.

Which species reactivity has this antibody been validated against?

Validated reactivity is listed on the PDP and on each lot's datasheet. We validate against the most commonly tested species for the target (pLenti-VGF-sgRNA); cross-reactivity in untested species may exist but is not guaranteed. Contact technical support for specific species inquiries.