pLenti-ACVR2A-sgRNA

Cat. No.: P743627

Specifications

Product Name

pLenti-ACVR2A-sgRNA

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

Item-specific, tested applications and recommended conditions: Not specified for this item; refer to CoA/Spec Sheet.

Because no validated protocols are provided in the Product Data, please consult institutional SOPs for:

• Plasmid preparation and transfection (if provided as DNA)
• Lentiviral packaging, titration, and BSL-2 handling (if provided as viral particles)
• Editing validation assays (amplicon NGS, ICE/TIDE, Western blot for ACVR2A/SMAD pathway readouts)

Any step-by-step procedures or dilution recommendations must be derived from the item’s official documentation when available.

Biological Roles

Item-specific facts (from Product Data):

• Target gene indicated by product name: ACVR2A.

Literature/general background on ACVR2A:

• ACVR2A (Activin A receptor type IIA) is a serine/threonine kinase receptor for activins within the TGF-β superfamily. Upon ligand binding (e.g., activin A), ACVR2A recruits and phosphorylates type I receptors (e.g., ACVR1B/ALK4), which then phosphorylate receptor-regulated SMADs (SMAD2/3). This leads to SMAD complex formation and transcriptional regulation affecting cell differentiation, proliferation, and endocrine signaling.
• ACVR2A participates in reproductive biology, development, and metabolic regulation pathways. It can form heteromeric receptor complexes and exhibits context-dependent signaling intensity shaped by ligand availability, co-receptor presence, and intracellular modulators (literature).
• Genetic perturbation of ACVR2A in model systems is used to dissect activin/TGF-β pathway wiring, feedback control (e.g., SMAD7), and crosstalk with MAPK/PI3K signaling.

Reagent context (general):

• An sgRNA construct targeting ACVR2A enables targeted knockout or transcriptional modulation (CRISPRi/a, when paired with dCas9 fusions). Validation should include checking on-target editing at ACVR2A exons, changes in SMAD2/3 phosphorylation following activin stimulation, and transcriptomic profiling of canonical activin-responsive genes.

Note: This section summarizes biological function for research context only and does not imply any clinical or diagnostic use.

Buffer Applications

This section is not typically applicable to a lentiviral sgRNA construct.

• Item-specific buffer composition and recommended dilution/working buffer are not specified for this item; refer to CoA/Spec Sheet.
• General note (literature/general): If provided as plasmid DNA, nuclease-free Tris/EDTA or similar low-salt buffers are commonly used for storage; for viral stocks, specialized cryostabilizing buffers are typical. Avoid repeated freeze–thaw cycles to maintain integrity.

Green Alternatives

Green chemistry substitution is not directly applicable to a nucleic acid/viral vector product in the same way it is for organic solvents or reagents. Nevertheless, sustainable practices can be applied to the workflows involving this product.

General recommendations (literature/general):

• Minimize single-use plastics by planning aliquots and using sterile, reusable components where permitted by biosafety.
• Scale appropriately to reduce reagent and consumable waste (optimize transfection/transduction at small scale before upscaling).
• Energy efficiency: Consolidate −80 °C storage and maintain freezers at optimal set points with good inventory control to reduce door-open time.
• Safer disinfectants: Use freshly prepared 10% bleach for viral inactivation, then neutralize and dispose per institutional guidelines; consider hydrogen peroxide–based surface disinfectants where compatible to lower chlorine footprint.
• Documentation: Keep accurate titer and activity records to avoid unnecessary repeat experiments.

Comparison (contextual):

• Delivery alternatives: Electroporation of RNPs (Cas9 protein + synthetic sgRNA) avoids viral packaging and associated biohazard waste, and can reduce off-target persistence. Trade-offs include transient activity and potentially lower efficiency in some cell types.
• Non-integrating vectors (e.g., IDLV, AAV for certain contexts) can lessen integration concerns but have different host ranges and capacity limits.

These are general considerations; adopt measures that align with your institutional biosafety and sustainability programs.

Pharmaceutical Uses

Not applicable. This product is labeled for research use only and is not intended for human or veterinary use.

General context (literature/general):

• Lentiviral sgRNA constructs are widely used in preclinical research to engineer cell lines, conduct genetic screens, and interrogate signaling pathways. They are not pharmaceutical excipients or drug substances. Any translational or manufacturing applications would require purpose-built GMP materials and regulatory oversight, which are outside the scope of this RUO product.

Physical Properties

Item-specific facts (from Product Data):

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration, buffer composition, A260/A280, endotoxin: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight/formula: Not specified for this item; refer to CoA/Spec Sheet.

General properties for plasmid/lentiviral DNA or lentiviral preparations (literature/general):

• Plasmid DNA behaves as a high–molecular weight polyanion in aqueous buffers; it does not have a meaningful boiling point/melting point like small molecules.
• If supplied as DNA in buffer, typical storage is frozen at −20 to −80 °C; repeated freeze–thaw can shear DNA. If supplied as lentiviral particles, viral stability is highest at −80 °C with single-use aliquots.
• Solubility: DNA is water-soluble (buffered aqueous media). Insoluble in nonpolar organic solvents. Osmolality, pH, and ionic strength depend on the buffer used (not specified here).
• Optical properties: Quantified by A260 for nucleic acid content; A260/A280 ~1.8–2.0 is typical for pure DNA (literature, not item-specific).

Note: Without a sequence map, backbone length, or formulation details, numerical specifications (e.g., size in bp, titer, refractive index, density) cannot be provided. Please consult the item’s CoA/Spec Sheet for exact values.

Quality and Grades

Item-specific facts (from Product Data):

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

General quality considerations for lentiviral sgRNA constructs (literature/general):

• Identity: Typically demonstrated by restriction digest pattern and/or full plasmid sequencing across the sgRNA insert and critical vector elements. For packaged virus, sgRNA cassette presence can be confirmed by qPCR of viral cDNA.
• Purity: For plasmid DNA, OD260/OD280 and OD260/OD230 ratios are used to assess protein and solvent carryover. Endotoxin level is critical if the plasmid is used to transfect sensitive cells or for in vivo research; “EndoFree” prep is often preferred.
• Titer (for lentiviral particles): Functional titer (TU/mL) or genome copies/mL by qPCR are common metrics. Stability is affected by storage and freeze–thaw history.
• Sterility: Mycoplasma- and bacteria-free status is important for cell culture work. Filtration through 0.22 µm and aseptic handling are standard.

Interpretation of grades (general):

• Research use only (RUO): Suitable for laboratory research; not cleared for diagnostic or therapeutic use.
• Sequence-verified: Confirms the intended guide and backbone (if provided). Absent here; please verify with the CoA/Spec Sheet or sequence map when available.

Reaction and Applications

Item-specific facts (from Product Data):

• Manufacturer Applications: Not specified for this item.

General research applications for pLenti-ACVR2A-sgRNA (literature/general):

• Genome editing: Guides CRISPR/Cas9-mediated cleavage at the ACVR2A locus to generate knockouts or targeted modifications when a donor template is provided. Cas9 must be supplied in trans (e.g., SpCas9-expressing cell line or co-delivery).
• Stable delivery: Lentiviral vectors enable stable integration in dividing and non-dividing cells, facilitating long-term expression of sgRNA for durable editing or CRISPRi/a when paired with dCas9-KRAB/VP64 systems.
• Pathway interrogation: ACVR2A encodes activin A receptor type IIA, a serine/threonine kinase receptor in the TGF-β/activin pathway. sgRNA-driven disruption supports studies of SMAD signaling, ligand responsiveness, and downstream transcriptional programs.
• Pooled screens: When incorporated into barcoded libraries, ACVR2A-targeting guides can participate in positive/negative selection screens to map pathway dependencies (library context, MOI control, and representation are critical).

Practical tips (general):

• Guide design and validation: Use on-target/off-target prediction tools; verify edits by amplicon NGS, T7E1, or ICE/TIDE. Clone-independent options (gBlocks/oligos) vary by backbone.
• Controls: Include non-targeting and safe-harbor (e.g., AAVS1) guides, plus rescue experiments with sgRNA-resistant cDNA where feasible.
• Editing outcomes: Expect indels via NHEJ; for HDR, synchronize cell cycle and supply donor DNA with appropriate homology arms.

Reaction Conditions

No chemical reaction conditions are applicable. Below are general, literature-based workflows for using a pLenti sgRNA construct; these are provided as non-item-specific guidance.

Typical lentiviral production (literature/general):

• Cells: HEK293T/293FT at ~70–90% confluence.
• Transfection: Co-transfect transfer vector (this sgRNA plasmid), packaging plasmids (e.g., psPAX2), and envelope plasmid (e.g., VSV-G) using a suitable transfection reagent. Change medium 6–18 h later.
• Harvest: Collect supernatant at 48 and 72 h, clarify by 0.45 µm filtration, concentrate if needed (PEG/ultracentrifugation). Determine titer (TU/mL) by functional assay or qPCR.

Typical transduction and editing (literature/general):

• Target cells: Choose permissive cells; for integration, maintain BSL-2 practices.
• MOI: 0.2–1 for pooled screens (to favor single integration), higher MOI for efficient editing in bulk populations.
• Selection: If the vector carries a selectable marker (not specified here), apply selection 48–72 h post-transduction.
• Cas9 provision: Use a stable Cas9 line or co-deliver Cas9. Validate on-target editing by amplicon sequencing or mismatch assays at 3–10 days.

Critical variables:

• Guide quality and genomic context; cell cycle; nuclease expression levels; and post-editing outgrowth time. Optimize empirically for your system. Refer to the item’s CoA/Spec Sheet for any vector-specific conditions if available.

Safety and Handling

Item-specific facts (from Product Data):

• GHS/CLP: Signal word, hazard statements, pictograms, classification: Not specified for this item; refer to SDS.
• Storage: Store at −80 °C.
• Shipping: Dry ice packs + Cold packs.

General guidance (literature/general):

• Biosafety: pLenti backbones are used to generate replication-incompetent lentiviral particles when co-transfected with packaging plasmids. The plasmid DNA itself is not infectious; however, any work producing or handling lentiviral particles should be performed under BSL-2 containment with institutional approvals and appropriate viral safety SOPs.
• PPE: Lab coat, gloves, and eye protection. Use a biological safety cabinet for procedures that may generate aerosols (e.g., pipetting viral stocks, vortexing, sonication of viral materials).
• Incompatibilities: Avoid DNases and microbial contamination. For lentiviral stocks, avoid repeated freeze–thaw; inactivate spills with freshly prepared 10% bleach (≥10–15 min contact) followed by appropriate cleanup.
• First aid (overview, defer to SDS): In case of skin/eye contact with buffers, rinse with water. If exposure to lentiviral particles occurs, follow institutional exposure protocols and report to EHS/Occupational Health.
• Waste: Treat materials contacting live lentiviral particles as biohazardous; decontaminate by bleach or autoclave per institutional policy.

Always consult the official SDS and institutional biosafety guidelines for authoritative, up-to-date safety information for this product’s exact format.

Solvent Selection

This product is a nucleic acid construct/viral reagent and is not used as a small-molecule solute in organic solvents.

• Item-specific: Solvent, buffer composition, and concentration are not specified for this item; refer to CoA/Spec Sheet.
• Practical note (general): Maintain aqueous, nuclease-free conditions. Avoid strong acids/bases and most organic solvents that denature DNA or disrupt viral envelopes. For plasmid DNA, TE or similar low-salt buffers are commonly used; for lentiviral stocks, specialized storage buffers with cryoprotectants may be employed (not specified here).

For tasks typically covered by “solvent selection” in chemistry, refer instead to the sections on Safety & Handling and Storage & Reconstitution for guidance relevant to biological reagents.

Storage and Reconstitution

Item-specific facts (from Product Data):

• Storage Conditions: Store at −80 °C.
• Shipped In: Dry ice packs + Cold packs.

Not specified for this item; refer to CoA/Spec Sheet for:

• Exact buffer formulation, concentration/titer, and presence of cryoprotectants
• Recommended aliquot size and thawing instructions
• Stability claims (shelf life at −80 °C, allowable freeze–thaw cycles)

General guidance (literature/general):

• Upon receipt, place immediately at −80 °C. If concentration/format permits, prepare single-use aliquots in sterile, nuclease-free tubes to minimize freeze–thaw events.
• Thaw on ice or at 4 °C. Mix gently by inversion; avoid vigorous vortexing, especially for lentiviral particles.
• For plasmid DNA, brief centrifugation after thaw ensures full recovery from tube walls; store working aliquots at −20 °C and archival stocks at −80 °C.
• Maintain aseptic technique to prevent nuclease and microbial contamination. Record lot, date, and number of freeze–thaw cycles for reproducibility.

Always follow any product-specific instructions supplied with the CoA/Spec Sheet.

Structure and Identity

Item-specific facts (from Product Data):

• Product: pLenti-ACVR2A-sgRNA (SKU: P743627)
• Format/Type: Lentiviral sgRNA construct targeting the ACVR2A gene (inferred from product name)
• CAS: Not applicable for a genetic construct
• SMILES / InChI / InChIKey: Not applicable for a nucleic acid construct
• Molecular formula, Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

General description (literature/general):

• This product name indicates a plasmid or lentiviral vector backbone engineered to express a single guide RNA (sgRNA) designed to target the ACVR2A locus for CRISPR-mediated genome editing. Such vectors typically drive sgRNA expression from a Pol III promoter (e.g., U6), while Cas9 is provided in trans or by a Cas9-expressing line. Specific promoter, selection markers, and sequence elements are not provided here.
• Structural features common to pLenti-style vectors (typical, literature): 5' and 3' LTRs (or SIN LTRs), packaging signal (Ψ), and common bacterial propagation elements (e.g., bacterial origin and antibiotic resistance marker). Exact features for this item are not specified.

2D/functional view in words (general):

• Linear functional map usually includes: LTR–Ψ–(sgRNA cassette)–LTR for the viral genome segment, plus a bacterial backbone for cloning/propagation. The sgRNA cassette encodes a short guide sequence (20 nt variable) fused to a scaffold/tracr sequence.

Synthetic Utility

Classical organic synthetic utility does not apply to a nucleic acid/viral vector. However, from a molecular biology standpoint, the construct has utility in assembling engineered cell systems.

General molecular biology utility (literature/general):

• Modular cloning: The sgRNA protospacer region (typically ~20 nt) can often be swapped by oligo ligation or Golden Gate cloning to retarget the vector. Multiple sgRNAs can be multiplexed using arrayed cassettes or independent promoters, depending on the backbone.
• Retrosynthetic design (functional): To achieve an ACVR2A loss-of-function phenotype, design exonic targets early in the coding sequence or at conserved kinase domains. For CRISPRi/a, target promoter-proximal regions (±200 bp of TSS) for maximal effect with dCas9-KRAB/activators.
• Integration with reporters: Pair with fluorescent or antibiotic selection markers (if present in the backbone; not specified here) to enrich edited populations before downstream assays.

Item-specific elements such as cloning sites, antibiotic resistance, or promoter identities are not provided here; consult the CoA/Spec Sheet or sequence map for exact design and cloning strategies.

Target Specificity

Item-specific facts (from Product Data):

• Intended gene target indicated by product name: ACVR2A (Activin A receptor, type IIA).

Not specified for this item; refer to CoA/Spec Sheet for the following details if available:

• Exact sgRNA protospacer sequence(s) and PAM context
• Species/isoform targeting (human/mouse/other orthologs)
• Off-target assessment or validation data
• Backbone details (promoter driving sgRNA, selectable markers)

General note (literature/general):

• Target specificity for CRISPR sgRNAs depends on perfect matching in the seed region adjacent to the PAM and chromatin accessibility at the target locus. Independent validation of edits and potential off-target sites is recommended in the chosen cell system.

Perguntas frequentes

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-ACVR2A-sgRNA); cross-reactivity in untested species may exist but is not guaranteed. Contact technical support for specific species inquiries.