pLenti-ARMC2-sgRNA

Cat. No.: P743901

Specifications

Product Name

pLenti-ARMC2-sgRNA

Product Line

Prodotti biologici

Stoccaggio e spedizione

Condizioni di conservazione di stoccaggio

Store at -80°C

Spedito in

Dry ice packs + Cold packs

Stabilità e conservazione

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

Application Protocols

Item-specific, tested applications

• Not provided for this item.

General protocol outline for using a pLenti sgRNA plasmid (literature/practice)

1. Design and verification

• Confirm sgRNA spacer sequence and PAM compatibility with the chosen Cas9. Sequence-verify the insert region.

2. Cell delivery

• For transient assays: Transfect plasmid DNA into cells expressing Cas9. For stable expression: Package into lentivirus and transduce target cells under BSL-2.

3. Selection/enrichment

• If the vector carries a selectable marker (not specified), apply selection (e.g., puromycin) or sort reporter-positive cells 48–72 h post-delivery.

4. Editing assessment

• Extract genomic DNA; PCR-amplify target locus; quantify editing by ICE/TIDE or NGS. Validate protein-level effects by immunoblot or flow cytometry.

5. Clonal isolation (optional)

• Single-cell clone by limiting dilution or FACS. Genotype clones to identify frameshift alleles.

6. Phenotypic assays

• Design readouts relevant to ARMC2 biology (e.g., cilia-related assays) as appropriate for the model system.

Notes

• Optimize conditions (DNA amount, MOI, selection stringency) per cell type. Maintain rigorous negative and positive controls.

Biological Roles

Item-specific target note

• Intended target gene (by product name): ARMC2. Specific sgRNA sequence and species targeting are not specified for this item; consult the CoA/Spec Sheet.

General/literature context on ARMC2

• ARMC2 (Armadillo Repeat-Containing protein 2) encodes a protein with armadillo repeat domains implicated in protein–protein interactions. Orthologs have been associated with ciliary/flagellar function and germ cell biology in several model organisms. Molecular roles may include participation in intraflagellar transport or structural scaffolding within motile cilia (literature). Exact functions can be cell-type and species dependent.

Use of ARMC2-targeting sgRNAs (general)

• Functional genomics: Disrupting ARMC2 via CRISPR can help interrogate its role in ciliogenesis, motility, or related signaling pathways in relevant cell models.
• Mechanistic studies: Pair sgRNA perturbation with transcriptomic or proteomic readouts to map downstream effects and interaction networks.

Notes

• No clinical or diagnostic claims are made. Researchers should verify species specificity, exon targeting, and potential isoform considerations before experiments.

Buffer Applications

Not typically applicable. This product is not a buffering agent.

General handling

• For reconstitution/dilution, nuclease-free water or TE buffer (10 mM Tris-Cl, 1 mM EDTA, pH 7.5–8.0) are commonly used for plasmid DNA (literature). Select buffer based on downstream application (e.g., avoid EDTA if divalent cations are required immediately after dilution).

Green Alternatives

This product is a biological DNA construct rather than a chemical solvent or reagent with conventional “green chemistry” substitutions.

General sustainability considerations (literature/practice)

• Minimize single-use plastics by consolidating aliquots and using reusable racks where biosafety allows.
• Prefer enzymatic cleanup/buffer exchange kits over organic-solvent precipitation when feasible to reduce solvent waste.
• Consolidate shipments (already shipped on dry ice + cold packs for this item) to reduce logistics footprint and dry-ice consumption.
• Where appropriate, consider non-viral delivery methods (electroporation or lipid nanoparticles) to avoid packaging workflows that generate additional consumable and disinfectant waste, acknowledging trade-offs in efficiency and cell-type compatibility.

Pharmaceutical Uses

Not applicable. This product is provided for research use only and is not intended for manufacturing or as an excipient.

General context (literature/practice)

• Lentiviral sgRNA plasmids are research tools for cell-based studies and process development. Any use in regulated manufacturing would require extensive characterization, GMP production, and regulatory review, which are beyond the scope of this catalog item.

Physical Properties

Item-specific properties

• Appearance/form: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration and buffer: Not specified for this item; refer to CoA/Spec Sheet.
• pH: Not specified for this item; refer to CoA/Spec Sheet.
• OD260/280: Not specified for this item; refer to CoA/Spec Sheet.

General/literature characteristics for plasmid DNA reagents

• Physical state: Aqueous solution or lyophilized DNA (varies by lot/formulation); plasmid DNA is a high–molecular weight, circular, double-stranded biopolymer.
• Solubility: Freely soluble in nuclease-free water or TE buffer (literature). Avoid divalent-cation–rich solutions if nucleases are a concern.
• Stability: Double-stranded plasmid DNA is stable at −20 to −80°C for long-term storage and at 4°C for short-term (days–weeks) when nuclease-free (literature). Minimize freeze–thaw cycles.
• UV characteristics: DNA absorbs at 260 nm; A260 of 1.0 ≈ 50 µg/mL for dsDNA (literature). This is general guidance; not a specification for this item.

Not applicable to macromolecular DNA

• Boiling/melting point, density, logP, and refractive index are not meaningful for plasmid DNA and are not provided.

Quality and Grades

Item-specific quality information

• Grade/purity: Not specified for this item; refer to CoA/Spec Sheet.
• QC attributes (e.g., sequence confirmation, endotoxin level, sterility): Not specified for this item; refer to CoA/Spec Sheet.

General context for plasmid DNA quality (literature/practice)

• Sequence identity: For sgRNA vectors, verification typically includes Sanger sequencing across the sgRNA insert and junctions, and backbone-unique features. Researchers should confirm sequence prior to critical experiments.
• Purity metrics: Common metrics include A260/280 and A260/230 ratios, supercoiled content by agarose gel, and absence of RNase/DNase contamination.
• Endotoxin: For transfection into sensitive cell lines or primary cells, low-endotoxin DNA is preferred. Endotoxin specifications vary by supplier; confirm on CoA if relevant.
• Sterility: DNA intended for cell culture should be produced and filled under sterile conditions and may be 0.22 µm–filtered.

Implications for use

• Higher purity and sequence-verified plasmid DNA generally yields improved transfection efficiency and more consistent gene editing outcomes. In the absence of item-specific grade, users should perform incoming QC suitable for their application (e.g., restriction digest, sequencing, test transfection).

Reaction and Applications

Item-specific manufacturer applications

• Not provided for this item.

General/literature applications for pLenti sgRNA vectors targeting ARMC2

• CRISPR-Cas9 gene perturbation: Delivery of an sgRNA targeting ARMC2 to enable knockout (with Cas9), CRISPRi (with dCas9-KRAB), or CRISPRa (with dCas9-activators), depending on the Cas9 context.
• Lentiviral delivery: pLenti backbones support packaging into replication-incompetent lentiviral particles for stable integration and long-term expression in dividing and non-dividing mammalian cells.
• Functional genomics: Use in loss-of-function studies, pathway interrogation, or validation screens where ARMC2 function is implicated.

Practical tips (general)

• Dry-run validation: Sequence the sgRNA insert and verify PAM compatibility with the Cas9 variant used (e.g., SpCas9 requires NGG PAM).
• Controls: Include non-targeting sgRNA and a positive control sgRNA (to an essential gene) to benchmark editing efficacy.
• Delivery: Optimize transfection/transduction conditions per cell type; titrate MOI for single-copy integration when desired. Polybrene can enhance transduction in many lines (check cytotoxicity).
• Selection and enrichment: If the backbone carries a selectable marker (not specified here), apply appropriate antibiotic or sort for fluorescent reporters. Validate editing via T7E1/ICE/TIDE or targeted amplicon NGS.
• Off-target mitigation: Use computational design and multiple independent sgRNAs to deconvolute on-target phenotypes.

Reaction Conditions

Not applicable to small-molecule reactions.

General workflows for use (literature/practice)

• Transfection: Optimize DNA amount (e.g., 0.5–2 µg per well of a 6-well plate) and reagent ratios per cell line protocol. Conditions vary widely; consult reagent technical notes.
• Lentiviral packaging: Co-transfect HEK293T-derived packaging cells with transfer vector, packaging plasmid(s), and envelope plasmid (e.g., VSV-G) following BSL-2 practices. Collect supernatant at 48–72 h. This is general guidance; not a specification for this item.
• Transduction: Add filtered viral supernatant with polybrene if tolerated; adjust MOI to control copy number. Spinoculation can improve efficiency in some lines.
• Editing window: Allow 48–96 h post-delivery before selection or analysis; full knockout phenotypes may require several days to weeks depending on protein turnover.
• Validation: Use PCR amplification across the target locus followed by T7E1/ICE/TIDE or amplicon sequencing; confirm protein loss by immunoblot if antibodies are available.

Safety and Handling

Item-specific safety data

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

General biosafety guidance for plasmid DNA and lentiviral vectors (literature/practice)

• Risk considerations: While purified plasmid DNA is typically not hazardous per GHS, vectors designed for lentiviral packaging are recombinant DNA materials. Handle under institutional biosafety approvals. Lentiviral particle generation and use commonly require BSL-2 practices.
• PPE: Laboratory coat, gloves, and eye protection. Use nuclease-free practices to maintain sample integrity.
• Engineering controls: Use a certified biological safety cabinet for manipulations involving lentiviral packaging, infected cells, or potentially aerosol-generating steps.
• Incompatibilities: Avoid nucleases (DNase), microbial contamination, and repeated freeze–thaw. Chemical incompatibilities are not generally applicable to aqueous DNA solutions, but avoid strong oxidizers or extremes of pH unless protocol requires and stability is verified.
• First aid (general): In case of skin/eye contact with solution, rinse with water. If inhalation exposure to aerosols of packaged lentivirus occurs, follow institutional exposure response. Always consult the SDS for authoritative guidance.
• Waste: Decontaminate surfaces and liquid waste with appropriate disinfectants (e.g., freshly prepared 10% bleach), and autoclave solid waste per institutional biosafety procedures.

Solvent Selection

Not typically applicable. This product is a plasmid DNA reagent, not a small-molecule solute selected for organic solvents.

General handling notes (literature/practice)

• Use nuclease-free water or TE buffer for dilution and storage. Avoid organic solvents which can denature DNA.
• If concentrating or exchanging buffers, use spin concentrators (nuclease-free, appropriate MWCO) or ethanol/isopropanol precipitation per standard molecular biology protocols.

Storage and Reconstitution

Item-specific instructions

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

General/literature guidance for plasmid DNA handling

• Upon receipt: Verify package integrity and transfer vials immediately to −80°C. If lyophilized, allow to equilibrate to room temperature in a desiccated environment before opening to avoid condensation.
• Reconstitution: If lyophilized, reconstitute with nuclease-free water or TE buffer to the desired concentration. Mix gently; avoid vigorous vortexing to preserve supercoiled topology.
• Aliquoting: Prepare single-use aliquots to minimize freeze–thaw cycles. Label with concentration, buffer, and date.
• Short-term storage: −20°C for weeks to months; 4°C for a few days if actively in use. Keep on ice during bench work.
• Avoid contamination: Use sterile, nuclease-free plastics and filter tips. Clean work area to minimize DNase exposure.

Notes

• Appearance, buffer composition, and concentration are not specified for this item; refer to the CoA/Spec Sheet and SDS for authoritative information.
• For research use only.

Structure and Identity

Item-specific facts (from Product Data)

• SKU: P743901
• Product name: pLenti-ARMC2-sgRNA
• CAS: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not applicable to plasmid DNA constructs.
• SMILES: Not applicable to macromolecular DNA constructs.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

General/literature description of construct type

• pLenti-ARMC2-sgRNA denotes a lentiviral plasmid backbone carrying a single-guide RNA (sgRNA) expression cassette designed to target the ARMC2 gene. Such vectors typically include: U6 or H1 promoter for sgRNA, tracrRNA scaffold, a lentiviral transfer backbone (e.g., 5′LTR–Ψ–RRE–cPPT–expression cassettes–3′LTR), and optional selectable markers (antibiotic resistance and/or fluorescent reporters). Specific elements for this item are not provided.
• 2D “structure” (verbal): A circular, double-stranded DNA plasmid comprising modular functional regions (promoters, RNA scaffold, selection marker, lentiviral packaging signals). No small-molecule structural formula applies.

Notes

• Sequence, backbone length, antibiotic resistance, and sgRNA target sequence are not specified for this item; consult the CoA/Spec Sheet or sequence map if provided by the supplier.

Synthetic Utility

Not applicable in the classical organic synthesis sense.

General molecular biology utility (literature/practice)

• Cloning flexibility: pLenti sgRNA vectors typically permit insertion or exchange of sgRNA spacers via Golden Gate or Gibson cloning strategies.
• Modular design: Backbones often allow swapping promoters, selection markers, or adding reporter cassettes, enabling tailored experimental design.
• Complementary constructs: Co-delivery with Cas9-expression plasmids or cell lines stably expressing Cas9 supports diverse CRISPR modalities (knockout, CRISPRi/a). Specific backbone compatibility for this item is not specified; consult sequence maps if available.

Target Specificity

Item-specific data

• Target name: ARMC2 (by product name).
• Species reactivity/target organism: Not specified for this item; refer to CoA/Spec Sheet.
• sgRNA spacer sequence/PAM context: Not specified for this item; refer to CoA/Spec Sheet.
• Off-target assessment/validation data: Not specified for this item; refer to CoA/Spec Sheet.

General notes (literature/practice)

• Specificity of CRISPR targeting depends on sgRNA sequence, genomic context, and nuclease variant. It is standard practice to design multiple independent sgRNAs and perform off-target prediction and empirical validation.