pLenti-ACTB-sgRNA

Cat. No.: P743597

Spécifications

Product Name

pLenti-ACTB-sgRNA

Product Line

Produits biologiques

Stockage et expédition

Conditions de stockage de stockage

Store at -80°C

Expédié en

Dry ice packs + Cold packs

Stabilité et stockage

Stored at -20 ℃, valid for 3 months;Stored at -80 ℃, valid for at least one year.

Application Protocols

Only information from product data may be listed here.

• Tested/validated applications (e.g., WB, IHC, IF, FC for antibodies; transfection/transduction parameters for vectors): Not specified for this item; refer to CoA/Spec Sheet.
• Recommended working dilutions, positive controls, or selection regimens: Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature)

• For plasmid DNA: Follow cell line–specific transfection optimization. Sequence-verify the insert prior to use.
• For lentiviral particles: Thaw on ice, keep cold, and avoid repeated freeze–thaw. Determine functional titer in your cell line and set MOI based on experimental goals. Employ appropriate biosafety measures.
• For CRISPR editing: Confirm Cas9 expression, include appropriate controls, and validate on-target outcomes by sequencing and phenotypic assays.

Biological Roles

Item-specific biological role

• Target specificity and guide sequence for this SKU are not specified; refer to CoA/Spec Sheet. The product name suggests association with ACTB, but definitive details are not provided in the Product Data.

General biology of ACTB (beta-actin) (literature)

• Function: Beta-actin is a highly conserved cytoskeletal protein involved in filament formation (F-actin), cell shape maintenance, motility, cytokinesis, intracellular transport, and mechanotransduction.
• Expression: ACTB is broadly and abundantly expressed in eukaryotic cells and is frequently used as a loading control in protein assays; however, expression can be context-dependent.
• Genetic considerations: Because ACTB contributes to essential cellular processes, complete loss-of-function may be lethal or strongly deleterious in many cell types. Heterogeneous editing outcomes and selection for hypomorphic alleles are common upon targeting essential genes.
• Network interactions: Actin dynamics are regulated by a wide array of actin-binding proteins (e.g., cofilin, profilin, ARP2/3 complex) and signaling pathways (Rho family GTPases). Perturbation of ACTB can have pleiotropic effects on cell physiology.

Implications for experiments (general)

• When employing sgRNA tools to perturb ACTB, consider time-resolved assays, inducible Cas9/dCas9 systems, or partial knockdown approaches to observe phenotypes while maintaining cell viability. Always validate on-target editing and evaluate off-target risks using appropriate controls and orthogonal assays.

Buffer Applications

Not typically applicable.

• This product is a genetic construct and is not used as a buffering agent or pH control component. Any buffer information pertinent to the storage or delivery of the construct (e.g., TE, PBS, proprietary media) is not specified for this item; refer to CoA/Spec Sheet.
• For experimental workflows, choose buffers that are nuclease-free and compatible with transfection/transduction reagents per standard molecular biology practice (general guidance).

Green Alternatives

Context

• This product is a biological tool (DNA/viral vector), not an organic solvent or commodity reagent. Green chemistry considerations focus on safer handling, reduced waste, and minimizing resource-intensive steps.

Greener practice options (general, literature)

• Delivery method choices: When feasible, non-viral delivery (e.g., plasmid transfection, RNP electroporation) can reduce biosafety burden compared with viral work, lowering decontamination needs and specialized containment.
• Process scale: Use high-titer, small-volume transductions to reduce plasticware and media waste. Plan experiments to minimize repeated freeze–thaw cycles (fewer, well-sized aliquots).
• Upstream production: If generating virus in-house, opt for serum-reduced media and optimized transient transfection to improve yield and reduce consumables per usable unit. Validate titer once to avoid redundant production.
• Downstream waste: Employ validated chemical disinfectants at minimal effective concentrations and ensure proper neutralization to limit corrosive waste. Autoclave only what is necessary to conserve energy.

Comparison snapshot (general)

• Viral delivery: High efficiency in difficult cells; requires BSL-2, generates disinfectant/autoclave waste.
• Non-viral (RNP/plasmid): Lower biosafety overhead; often lower integration risk; may need repeat dosing; typically less hazardous waste.

Item-specific greener attributes

• Not specified for this item; refer to CoA/Spec Sheet for formulation details that may impact waste handling (e.g., presence of serum or cryoprotectants).

Pharmaceutical Uses

Not applicable to this catalog item.

• This product is intended for research use only and is not an excipient, API, or GMP-manufactured material. No pharmacopeial status, formulation roles, or clinical manufacturing uses are specified or implied for this SKU.
• If translating research findings, separate sourcing and qualification under appropriate quality systems would be required; this note is general context only and not specific to this item.

Physical Properties

Item-specific properties

• Concentration, buffer composition, pH, and excipients: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight, formula, and small-molecule physicochemical data (BP, MP, density, logP, refractive index): Not applicable to DNA/viral constructs; not specified for this item.

General properties for nucleic acid constructs (literature)

• Physical form: Typically provided as an aqueous solution or frozen aliquots (e.g., TE or proprietary buffer) for plasmid DNA, or frozen viral supernatants/partially purified preparations for lentiviral particles. Exact formulation for this SKU is not specified.
• Solubility: DNA is water-soluble; viral particles are suspended in aqueous media; avoid repeated freeze–thaw to maintain integrity/titer (general guidance).
• Stability: sgRNA expression cassettes in plasmids are stable when stored at -20 to -80°C; lentiviral particles are commonly stored at -80°C to minimize titer loss. This product specifies -80°C storage.
• Optical properties: Nucleic acids exhibit UV absorbance with A260 used for quantification; A260/A280 is a common purity metric for plasmids (literature practice). Not specified for this item.
• Viscosity/osmolality: Dependent on buffer and concentration; not specified for this item.

Notes

• Because this is a biological construct rather than a defined small molecule, conventional physicochemical constants (e.g., boiling point) do not apply. For lot-specific details (vector length in bp, exact buffer, titer if viral), consult the CoA/Spec Sheet.

Quality and Grades

Item-specific grade/purity

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

General expectations for CRISPR sgRNA lentiviral/plasmid reagents (literature/typical)

• Plasmid DNA preparations for transfection are commonly endotoxin-reduced and sequence-verified. For viral formats, typical quality attributes include sterility, mycoplasma negativity, and functional titering (e.g., TU/mL). None of these attributes are specified here; consult the CoA for lot-specific results.
• Sequence verification: For sgRNA constructs, Sanger confirmation of the guide spacer and junctions is standard. Backbone identity is typically confirmed by restriction analysis or NGS. Not specified for this item.
• Stabilizers: Viral preparations sometimes include serum proteins, sugars, or cryoprotectants to improve freeze–thaw stability; plasmids are often provided in TE or similar buffers. Any stabilizers for this SKU are not specified.

How to use quality information

• Use the CoA/Spec Sheet to verify: vector map and features (promoters, selection marker, reporter), sgRNA spacer sequence, plasmid length, antibiotic resistance, and—if viral—the functional titer and sterility testing. These determine dosing (e.g., MOI), selection regimen, and biosafety level.
• If performing critical experiments (e.g., essential gene knockout), independently verify the sgRNA sequence and, if viral, re-titer under your cell-line conditions to calibrate MOI and selection endpoints.

Reaction and Applications

Item-specific applications (from catalog)

• Manufacturer Applications: Not specified for this item.

General applications for pLenti sgRNA constructs (literature/typical)

• Gene knockout (CRISPR-Cas9): sgRNA directs Cas9 to the ACTB locus to induce DSBs and NHEJ-mediated indels, generating loss-of-function alleles. Requires co-expression of Cas9 (stable or transient). The exact guide sequence and PAM compatibility for this SKU are not specified.
• Transcriptional modulation: When paired with dCas9-based effectors, the sgRNA can enable CRISPRi (repression) or CRISPRa (activation) of ACTB. Efficacy depends on target site position and effector architecture.
• Pooled screening: Lentiviral delivery allows stable integration for pooled functional genomics; an ACTB-targeting guide can serve as a positive control for essential gene targeting in some contexts. Use appropriate biosafety and MOI control.
• Marker-based enrichment: Many sgRNA vectors include selectable markers (e.g., antibiotic resistance or fluorescent reporters) to enrich edited populations. Specific markers for this item are not provided; refer to CoA/Spec Sheet.

Practical tips (general)

• Validate on-target activity with sequencing (amplicon NGS or TIDE/ICE) and confirm protein-level effects (e.g., Western blot for beta-actin) when biologically appropriate. For essential genes, anticipate strong selection against complete knockout; titrate conditions accordingly.
• Include non-targeting and safe-harbor guides as controls. Assess potential off-targets using in silico tools and empirical assays where critical.

Reaction Conditions

Item-specific conditions

• Recommended transfection/transduction conditions, cell types, MOI, antibiotic selection, and exposure times: Not specified for this item; refer to CoA/Spec Sheet.

General conditions for sgRNA delivery (literature)

• Plasmid transfection: Optimize DNA amount and transfection reagent ratio per cell line; verify Cas9 availability (stable line or co-transfection). Selection/enrichment depends on vector marker; follow the marker’s standard regimen.
• Lentiviral transduction: Determine functional titer and choose a multiplicity of infection (MOI) appropriate for your application (e.g., MOI ≈0.3–1 for single-copy integration in pooled settings; higher MOI for saturation in arrayed assays). Enhance transduction with polycations where appropriate and permissible. Replace media after 12–24 h and begin selection per marker guidance, if present.
• Timeline: Indel formation can be detected within 48–96 h after delivery of Cas9+sgRNA; clonal isolation and expansion require 1–3 weeks depending on cell growth.
• Readouts: Use amplicon sequencing or mismatch-detection assays to quantify editing; confirm protein-level changes by immunoblotting or functional assays relevant to actin biology.

Controls and QC

• Include non-targeting and safe-harbor control guides. Monitor off-target activity using predictive tools and orthogonal assays where outcomes are critical.
• Always adapt conditions to the specific vector features provided in the CoA/Spec Sheet for this SKU.

Safety and Handling

Item-specific hazard information

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

General laboratory safety (defer to institutional biosafety policies and SDS)

• Personal protective equipment: Wear lab coat, gloves, and eye protection. Work in a certified biological safety cabinet (Class II) if handling lentiviral particles or large volumes of DNA to prevent aerosol exposure (general guidance).
• Biosafety level: Lentiviral vector work is typically conducted at BSL-2 with appropriate approvals and training; follow local IBC guidance. If the item is a plasmid DNA only, standard BSL-1/BSL-2 practices may apply depending on downstream use. The specific format for this item is not specified; consult SDS/CoA and your IBC.
• Incompatibilities and precautions: Avoid generating aerosols; disinfect spills with appropriate virucidal/biocidal agents (e.g., freshly prepared 10% bleach followed by 70% ethanol) when working with viral material. Use nuclease-free practices to avoid degradation of nucleic acid constructs.
• First aid (general overview): In case of skin/eye contact with solution, rinse with water for several minutes; remove contaminated clothing. If inhalation of aerosols is suspected, move to fresh air and seek medical evaluation per institutional protocols. Ingestion is not an intended route; seek medical attention if exposure occurs. Always defer to SDS for authoritative measures.
• Waste: Decontaminate biological waste by chemical disinfectant and/or autoclaving according to institutional biosafety guidelines.

Special risks

• Replication-incompetent lentiviral systems can integrate cargo into host genomes; take appropriate containment measures. If handling plasmid DNA, risk is primarily low but depends on downstream use. Details for this SKU are not specified; refer to SDS/CoA.

Solvent Selection

This product is a biological construct (nucleic acid and/or viral suspension), not a low–molecular-weight solute; classical solvent selection is not typically applicable.

General handling guidance (literature)

• Aqueous compatibility: DNA constructs and viral suspensions are maintained in aqueous buffers. Use nuclease-free water or suitable molecular biology buffers when diluting or formulating. Avoid organic solvents that destabilize proteins/envelopes (for viral preps) or that can precipitate DNA (e.g., ethanol/isopropanol) unless performing intentional precipitation/purification steps.
• Additives: If concentrating or aliquoting viral supernatants, maintain isotonic conditions and consider cryoprotectants as appropriate per institutional SOPs. Exact formulation for this SKU is not specified; refer to CoA/Spec Sheet.

When to consider alternatives

• If solvent compatibility is a concern for a downstream assay (e.g., small-molecule additions), mix components after transduction/transfection to minimize exposure of the biological reagent to organic co-solvents. For DNA-only transfections, short exposures to common transfection mixes are usually tolerated (per reagent manufacturer).

Storage and Reconstitution

Item-specific instructions (from Product Data)

• Storage conditions: Store at -80°C.
• Shipped in: Dry ice packs + Cold packs.

General handling for this class of products (literature)

• Thawing: Thaw on ice or at 4°C. Mix gently by pipetting; avoid vigorous vortexing to preserve integrity (especially for viral particles).
• Aliquoting: Upon first thaw, aliquot into single-use volumes to minimize freeze–thaw cycles, which can reduce viral titer or nick plasmid DNA. Use sterile, nuclease-free tubes and tips.
• Short-term storage: Keep on ice during setup. If repeated use over a few days is necessary, store at 4°C only if recommended in the CoA; otherwise, re-freeze promptly at -80°C.
• Long-term storage: Maintain at -80°C as specified. Avoid frost-free freezers and temperature fluctuations. Consider secondary containment to prevent desiccation.
• Reconstitution/dilution: If provided lyophilized (not specified for this item), reconstitute with nuclease-free water or supplied buffer per datasheet. For liquid formulations, dilute with sterile, nuclease-free buffer suitable for your application (e.g., cell culture medium for viral transduction) following institutional biosafety protocols.
• Documentation: Record lot number, thaw/aliquot dates, and aliquot locations to track usage and mitigate activity loss.

Note

• Exact buffer composition, concentration, and any stabilizers are not specified for this item; refer to the CoA/Spec Sheet for format-specific guidance.

Structure and Identity

Item-specific facts (from Product Data)

• SKU: P743597
• Product name: pLenti-ACTB-sgRNA
• Storage: Store at -80°C
• Shipping: Dry ice packs + Cold packs
• Research use: For research use only
• CAS/CID/SMILES/InChIKey/Molecular formula/Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

General description (literature/typical for this class)

• pLenti-ACTB-sgRNA denotes a lentiviral vector backbone (“pLenti”) carrying a single guide RNA (sgRNA) designed to target the ACTB locus (beta-actin) for CRISPR-based genome editing or modulation. Exact sequence, promoter(s), selection marker(s), and backbone map are not specified for this item.
• Structural features (typical for lentiviral sgRNA vectors): U6 or H1 RNA polymerase III promoter driving a 20-nt spacer within an invariant sgRNA scaffold; lentiviral LTRs and packaging signals (ψ), plus common accessory elements (e.g., RRE, cPPT/CTS) enabling packaging into VSV-G–pseudotyped particles when co-transfected with helper plasmids. These features are general and may vary; consult the product CoA/Spec Sheet for the definitive map for this SKU.
• Molecular entity type: engineered nucleic acid construct (double-stranded DNA plasmid backbone for production of lentiviral particles, or a prepared lentiviral particle suspension). The specific format for this item is not specified; refer to CoA/Spec Sheet.

2D description in words (general)

• The construct comprises a lentiviral backbone with terminal LTRs surrounding a cassette that includes an RNA polymerase III promoter linked to an sgRNA scaffold containing an ACTB-targeting spacer sequence. Additional vector elements (selection marker, reporter, origin of replication) are vector-dependent and not specified for this item.

Synthetic Utility

This section is interpreted for molecular biology “construction” utility rather than organic synthesis.

General utility of pLenti sgRNA vectors (literature)

• Modular cloning: sgRNA spacers are commonly introduced by oligo annealing and ligation, Golden Gate assembly (e.g., BsmBI/BbsI sites), or Gibson/NEBuilder methods. The exact cloning sites/backbone for this SKU are not specified.
• Payload exchange: Lentiviral backbones frequently support swapping promoters, reporters, and selection cassettes, enabling tailored gene-editing workflows (e.g., co-expression of Cas9, fluorescent markers). Vector map and restriction schema for this item are not provided; refer to CoA/Spec Sheet.
• Combinatorial perturbations: Multiple sgRNA expression units can be assembled (polycistronic cassettes or multiplex vectors) for targeting several loci. Efficiency depends on promoter choice and spacer design.
• Interoperability: Many labs use standardized U6-driven sgRNA scaffolds compatible with S. pyogenes Cas9 (SpCas9, PAM NGG) or alternative nucleases (SaCas9, AsCas12a, etc.). Compatibility for this item is not specified.

Practical notes

• Verify final constructs by Sanger sequencing across the promoter–sgRNA junction and the spacer region. Maintain plasmids in recombination-deficient strains to preserve repeats and complex elements (general advice). If using viral particles, confirm integration by PCR of genomic DNA post-transduction.

Target Specificity

Only information from product data may be listed here.

• Antigen/target name: Not specified for this item; refer to CoA/Spec Sheet.
• Species reactivity: Not specified for this item; refer to CoA/Spec Sheet.
• Clone/isotype (if antibody) or sgRNA spacer sequence/PAM context (if nucleic acid tool): Not specified for this item; refer to CoA/Spec Sheet.
• Validated applications: Not specified for this item; refer to CoA/Spec Sheet.

Note: The product name suggests relevance to ACTB, but definitive target sequence and specificity parameters for this SKU are not provided in the Product Data.