pLenti-ARHGAP9-sgRNA

Cat. No.: P743876

Specifications

Product Name

pLenti-ARHGAP9-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

Only product-specific, tested applications and conditions should be listed here; none are provided in the Product Data for this item.

• Tested applications: Not specified for this item; refer to CoA/Spec Sheet.
• Recommended conditions (MOI, selection antibiotics, transfection reagents, titration methods): Not specified for this item; refer to CoA/Spec Sheet.

General suggestion

• Follow standard institutional protocols for cloning verification, lentiviral packaging, transduction, and genome editing assessment appropriate to your cell model and biosafety level.

Biological Roles

Scope

• This tab addresses biology of the nominal target gene ARHGAP9 in general literature terms. It does not describe specific performance of this product.

ARHGAP9 overview (literature)

• ARHGAP9 encodes a Rho GTPase-activating protein (RhoGAP) that enhances the intrinsic GTPase activity of Rho-family small GTPases, promoting the GDP-bound (inactive) state. RhoGAPs regulate actin cytoskeleton dynamics, cell adhesion, polarity, and migration.
• Reported contexts include modulation of signaling pathways linked to cytoskeletal remodeling and possibly MAPK pathway crosstalk. Tissue and cell-type expression patterns vary; isoform complexity has been noted for several RhoGAPs. Specific ARHGAP9 roles may be cell-context dependent.

Implications for research (literature)

• Genetic perturbation of ARHGAP9 (knockout/knockdown or CRISPRi/a) is commonly used to probe cytoskeletal regulation, motility, and signaling networks. Phenotypic assays may include changes in stress fiber formation, focal adhesion markers, and migration/invasion readouts.

Item-specific statements

• The Product Data do not specify guide sequence, species targeting, or validation results for ARHGAP9 editing. Users should confirm genome compatibility, off-target risk, and functional outcomes experimentally for their system.

Buffer Applications

Not typically applicable.

• This product is a plasmid/viral vector reagent and is not used as a buffering agent. Handle in nuclease-free aqueous buffers appropriate for molecular biology (e.g., TE, PBS for formulated viral stocks). Any buffer composition for this specific item is not specified; refer to the CoA/Spec Sheet if provided.

Green Alternatives

This product is a biological plasmid/viral vector reagent rather than a chemical solvent or reagent traditionally assessed by green metrics (e.g., volatility, toxicity, lifecycle carbon intensity).

• Not typically applicable: There are no direct “green solvent” substitutions.
• Sustainability considerations (general):
  • Minimize single-use plastics by consolidating aliquots where biosafety allows.
  • Use room-temperature-stable enzymes/kits when feasible to reduce cold-chain burden (not applicable to this item’s storage requirement, which is −80°C as specified).
  • Plan experiments to reduce repeat freeze–thaw cycles and waste of packaged viral stocks.

Item-specific environmental data (biodegradability, solvent alternatives, energy use): Not specified for this item; refer to institutional sustainability guidelines.

Pharmaceutical Uses

Not applicable.

• Research Use Only: Per Product Data, this item is for research use only and is not intended for diagnostic procedures, clinical applications, or as a pharmaceutical excipient or active.
• No pharmacopeial status, formulation role, or cGMP information is provided. For translational work, obtain materials with appropriate regulatory documentation and quality attributes.

Physical Properties

Item-specific facts (from Product Data)

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

General properties for plasmid DNA reagents (literature)

• Solubility: Highly water-soluble in standard molecular biology buffers (e.g., TE, nuclease-free water). Insoluble in most organic solvents.
• Stability: Sensitive to nucleases and repeated freeze–thaw. Long-term stability improved by storage at −80°C (item-specified) in low-salt, EDTA-containing buffers to chelate divalent cations.
• UV characteristics: DNA exhibits λmax ~260 nm (literature); A260 of 1.0 ≈ 50 µg/mL for dsDNA (literature). Not a product specification.
• pH considerations: Common handling buffers pH 7.5–8.5 (literature); specific pH for this item is not specified.

Important

• No melting point, boiling point, density, refractive index, logP, or pKa are applicable for plasmid DNA as a formulated aqueous biomolecular reagent.
• For any lot-specific quality metrics (size confirmation, sequence verification, endotoxin, residual solvents), consult the item’s CoA/Spec Sheet (not provided here).

Quality and Grades

Item-specific facts (from Product Data)

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

Interpretive guidance (general for plasmid/viral tools)

• Research Use Only (RUO): Intended for laboratory research. Not for diagnostic, therapeutic, or human/animal administration.
• Typical QC for CRISPR sgRNA vectors (literature): Sequence verification of guide and scaffold; restriction digest or electrophoretic confirmation of backbone; optional endotoxin assessment for cell culture; sterility testing for packaged viral stocks. These are general practices; this item’s specific QC acceptance criteria are not provided.
• UV/trace impurities: Chromatographic or spectrophotometric metrics (A260/280, A260/230) are often used to assess purity of plasmid preps but are not specified here.
• Functional qualification: Users often validate cutting efficiency by T7E1/Surveyor assays, Sanger ICE/TIDE analysis, or NGS. No functional data are provided for this item.

Recommendation

• Review the lot-specific CoA/Spec Sheet for exact grade, sequence confirmation, backbone map, antibiotic marker, and any endotoxin/specification limits (not included in the Product Data).

Reaction and Applications

Item-specific facts (from Product Data)

• Application text: Not provided.

General applications for pLenti sgRNA vectors (literature)

• Gene perturbation: Expression of an sgRNA targeting ARHGAP9 to support CRISPR/Cas9-mediated knockout, repression (CRISPRi; with dCas9-KRAB), or activation (CRISPRa; with dCas9-VP64/p300) depending on the Cas architecture supplied by the user. The actual guide sequence, PAM, and orientation for this item are not specified.
• Lentiviral delivery: pLenti backbones enable stable integration (when packaged with helper plasmids) for long-term expression in dividing and non-dividing cells. Use appropriate BSL-2 controls and integration-aware experimental design.
• Functional genomics: Suitable for single-gene interrogation, pooled screening (if format-compatible), or pathway analysis related to Rho GTPase signaling mediated by ARHGAP9 (general biology; not product-validated).

Practical tips (general)

• Verification: Confirm the sgRNA insert and junctions by Sanger sequencing using vector-specific primers prior to use.
• Packaging workflow: Co-transfect HEK293T or similar with packaging (gag/pol), envelope (e.g., VSV-G), and the pLenti-sgRNA vector; harvest supernatant at 48–72 h. Titer and transduce target cells at a suitable MOI.
• On-target assessment: Evaluate editing with mismatch detection assays, ICE/TIDE, or deep sequencing; confirm loss-of-function at protein level when applicable.

Note: No item-specific efficiencies, titers, or markers are provided in the Product Data.

Reaction Conditions

Item-specific conditions: Not specified for this item; refer to CoA/Spec Sheet.

General literature guidance for use of pLenti sgRNA vectors

• Bacterial propagation: Transform into a recombination-deficient E. coli strain; culture at 30–37°C with appropriate antibiotic. Avoid overgrowth to reduce recombination of lentiviral LTRs.
• Plasmid preparation: Use endotoxin-reducing kits for sensitive cell types. Elute in nuclease-free buffer (e.g., 10 mM Tris, 0.1–1 mM EDTA, pH ~8) for storage.
• Transfection (packaging cells): HEK293T or derivatives; DNA:transfection-reagent ratios per manufacturer (e.g., PEI, lipids). Typical incubation at 37°C, 5% CO2; harvest viral supernatants 48–72 h post-transfection. Filter (0.45 µm) and, if needed, concentrate by ultracentrifugation or PEG precipitation.
• Transduction: Choose MOI to balance single-copy integration versus efficiency (commonly MOI ~0.3–1 for clonal KO work; higher for bulk edits—literature guidance). Include polybrene if compatible with cells. Select with antibiotic when backbone provides resistance (marker for this item is not specified).
• Editing assessment: 48–96 h post-transduction for initial genotyping; longer for protein turnover-dependent phenotypes. Use ICE/TIDE/NGS for indel spectra.

All parameters above are general guidance only; optimize per cell line and workflow. Follow institutional biosafety protocols for all lentiviral work.

Safety and Handling

Item-specific facts (from Product Data)

• Storage conditions: Store at −80°C.
• Hazard information (Signal word, H-statements, GHS class, pictograms): Not specified for this item; refer to SDS.
• Research use note: For research use only.

General guidance for plasmid CRISPR vectors (literature/best practice; defer to SDS and IBC policies)

• Biosafety: Plasmid DNA itself is not replication-competent or infectious; however, when used to produce lentiviral particles, work should be conducted under BSL-2/2+ conditions per institutional biosafety protocols. Avoid generating aerosols. Use certified biosafety cabinets for packaging/transduction.
• PPE: Laboratory coat, gloves, and eye protection. Change gloves frequently to minimize nuclease contamination.
• Handling precautions: Maintain nuclease-free technique. Keep on ice during handling. Avoid repeated freeze–thaw cycles; aliquot upon receipt.
• Incompatibilities: Avoid exposure to nucleases (DNase), high temperatures, and divalent cations without chelators (can promote nuclease activity). Avoid strong oxidants/UV that may damage DNA.
• First aid (overview): In case of accidental exposure to packaging mixtures or waste, follow institutional protocols; wash affected skin with water and seek medical attention per SDS guidance.
• Waste: Deactivate lentiviral supernatants with appropriate disinfectants (e.g., freshly prepared bleach) before disposal per local regulations. Plasmid waste may be autoclaved where permitted.

Always consult the SDS and your institutional biosafety committee (IBC) for authoritative safety requirements.

Solvent Selection

This product is a nucleic acid reagent and is not used as a reaction solvent.

• Item-specific solvent details: Not specified for this item; refer to CoA/Spec Sheet.
• Practical note: Handle in nuclease-free aqueous buffers (e.g., TE or nuclease-free water). Avoid organic solvents which can denature or precipitate DNA. For cell culture or transfection, dilute into appropriate sterile, buffered media as per your protocol.

For solvent considerations related to chemical synthesis, refer instead to the Reaction & Applications and Reaction Conditions tabs for biology-focused handling guidance.

Storage and Reconstitution

Item-specific facts (from Product Data)

• Storage conditions: Store at −80°C.
• Shipping: Dry ice packs + cold packs.
• Formulation/concentration/reconstitution: Not specified for this item; refer to CoA/Spec Sheet.

General guidance for plasmid/viral vector reagents (literature)

• Aliquoting: Upon receipt, thaw on ice, mix gently, and prepare single-use aliquots to avoid freeze–thaw cycles that can degrade DNA or reduce viral titer.
• Short-term handling: Keep on ice during setup. For plasmid DNA, brief storage at 4°C (hours to a few days) is acceptable; return to −20/−80°C for longer terms. For viral stocks, maintain at 4°C only for very short periods (≤24–48 h) and store at −80°C for longer.
• Thawing: Thaw rapidly on ice or at 4°C. Do not vortex viral stocks; invert gently.
• Buffer considerations: Nuclease-free TE (10 mM Tris, 0.1–1 mM EDTA, pH ~8) can enhance DNA stability. Exact buffer for this item is not specified.
• Documentation: Retain lot number and any sequence/map files in your LIMS for traceability.

Always defer to the CoA/Spec Sheet for lot-specific storage, formulation, and reconstitution instructions.

Structure and Identity

Item-specific facts (from Product Data)

• Product name: pLenti-ARHGAP9-sgRNA (SKU: P743876)
• CAS/CID/SMILES/InChIKey: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula / Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

General description (literature/typical for pLenti CRISPR-sgRNA constructs)

• Construct type: Lentiviral CRISPR single-guide RNA (sgRNA) expression vector designed to target the ARHGAP9 gene. Actual guide sequence, PAM context, and backbone elements are not specified for this item.
• Typical backbone features (literature): 5′ and 3′ LTRs, Ψ packaging signal, RRE, cPPT, and a Pol III promoter (commonly U6 or H1) driving the sgRNA scaffold; many pLenti CRISPR backbones also include a selection marker (e.g., puro or neo) and sometimes a reporter (e.g., GFP). Presence/identity of these features for this item is not specified.
• 2D “structure” in words: Double-stranded circular DNA plasmid containing lentiviral long terminal repeats flanking an sgRNA expression cassette targeted to ARHGAP9; additional vector modules may include origin(s) of replication and antibiotic resistance for propagation in E. coli (typical for such vectors; not item-specified).

Notes

• This listing represents a molecular biology reagent (plasmid/viral vector component), not a small-molecule chemical; therefore, conventional chemical identifiers (SMILES, InChIKey) are generally not applicable.

Synthetic Utility

Context

• For a molecular biology vector, “synthetic utility” maps to cloning and construct engineering rather than chemical synthesis.

General cloning utility (literature)

• sgRNA cassette engineering: Many pLenti backbones support insertion of custom protospacers via Type IIS Golden Gate cloning (e.g., BsmBI/BbsI) or Gibson/SLiCE assembly. Overhangs and cloning sites vary by vector design; specifics for this item are not provided.
• Modular assembly: Lentiviral backbones often permit exchange of promoters, selection markers, or reporters through unique restriction sites or sequence-homology assembly. This enables construction of multi-guide arrays, inducible sgRNA expression, or combinatorial libraries.
• Rescue/orthogonal studies: Pair ARHGAP9 sgRNA with cDNA rescue constructs (silenced to sgRNA by synonymous mutations) to validate on-target phenotypes.

Practical notes

• Verify plasmid maps and restriction site compatibility prior to assembly. Sequence all junctions. Maintain high-fidelity polymerases for any PCR-amplified fragments.

Item-specific constraints

• Backbone map, antibiotic resistance, cloning sites, and guide insertion method are not specified for this item; consult the CoA/Spec Sheet or vendor documentation.

Target Specificity

Item-specific facts (from Product Data)

• Target designation inferred from product name: ARHGAP9. No additional details (guide sequence, species genome, PAM context, off-target analysis) are provided.
• Clone ID, vector backbone, selectable marker, reporter, and insert verification: Not specified for this item; refer to CoA/Spec Sheet.

Notes

• Because this is an sgRNA construct (not an antibody), conventional antibody-specific attributes (epitope, clone/isotype, species reactivity) do not apply.
• Users should confirm genome build, target exon(s), and potential off-target sites experimentally for their system.