pLenti-YTHDF2-sgRNA

Cat. No.: P751896

Especificaciones

Product Name

pLenti-YTHDF2-sgRNA

Product Line

Productos biológicos

Almacenamiento y envío

Condiciones de almacenamiento de almacenamiento

Store at -80°C

Enviado en

Dry ice packs + Cold packs

Estabilidad y almacenamiento

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

Application Protocols

Item-specific tested applications, protocols, and recommended conditions are not provided in the Product Data for this item.

Please refer to the CoA/Spec Sheet for any lot-specific protocols. General workflow suggestions can be found under Reaction Conditions, but these are literature-based and not validated for this specific product.

Biological Roles

Item-specific facts (from Product Data)

• The construct name indicates targeting of YTHDF2; no further biological details are specified.

General background on the target (literature/general)

• YTHDF2 is a cytoplasmic m6A reader protein that binds N6-methyladenosine-modified mRNAs and promotes their decay by recruiting deadenylation/decapping machinery, thereby shaping transcript stability and gene expression programs.
• Functional domains include the C-terminal YTH domain that recognizes RRACH consensus m6A marks and N-terminal low-complexity regions implicated in RNA/protein interactions.
• Genetic perturbation of YTHDF2 is widely used to probe: mRNA turnover kinetics, stress responses, developmental regulation, and RNA modification networks. sgRNAs against YTHDF2 help delineate direct vs indirect effects of m6A-mediated pathways in cultured cells and model systems.

Note

• The biological summary above is provided for research context only. This product is for research use, not for diagnostic or therapeutic applications. Item-specific target isoforms/species and guide sequence are not specified; consult the CoA/Spec Sheet if provided.

Buffer Applications

Not typically applicable as a distinct buffer system topic for plasmid DNA.

• Working solutions are commonly prepared in nuclease-free water or TE buffer (10 mM Tris-Cl, 1 mM EDTA, pH ~8.0) to stabilize DNA and chelate divalent cations that catalyze nuclease activity (literature/general). Specific formulation for this item is not specified; refer to CoA/Spec Sheet.
• For cell culture or transfection, follow reagent manufacturer’s recommended buffers and opti-media; these are application-dependent rather than intrinsic to the plasmid.

Green Alternatives

This section is not typically applicable to plasmid DNA reagents.

• Environmental impact considerations primarily involve minimizing single-use plastics, optimizing transfection to reduce repeats, and proper neutralization of bleach before disposal.
• For chemical solvent substitution or greener solvent choices, see sections relevant to small-molecule reagents; not applicable here.

Pharmaceutical Uses

This product is supplied for research use only; no pharmacopeial status or clinical/formulation use is claimed or implied.

General notes (literature/general)

• CRISPR plasmids and sgRNA constructs can serve as process development tools in preclinical research and cell line engineering. Any use in GMP or clinical environments requires extensive additional qualification, documentation, and regulatory review distinct from research-grade materials.
• For translational workflows, typical requirements include: sequence confirmation, sterility testing, low endotoxin specifications, residual host contaminant testing, and release assays—none of which are specified for this item in the provided data.

Physical Properties

Item-specific facts (from Product Data)

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration, form (lyophilized/solution), A260/A280, endotoxin: Not specified for this item; refer to CoA/Spec Sheet.

General properties for plasmid DNA (literature/general)

• Physical state: circular double-stranded DNA; commonly supplied as aqueous solution or lyophilized solid.
• Solubility: highly soluble in nuclease-free water and TE buffer; insoluble in most organic solvents.
• UV absorbance: strong at 260 nm; typical A260/A280 ~1.8–2.0 for pure DNA (literature).
• Stability: stable at −20 to −80°C for long-term storage; susceptible to nucleases and repeated freeze–thaw.
• pH sensitivity: optimal stability near neutral pH; depurination accelerates under acidic conditions (literature).

Not applicable to nucleic acid macromolecules

• Boiling point, melting point (thermodynamic), logP, refractive index: not meaningful for plasmid DNA; not specified for this item; refer to CoA/Spec Sheet if provided for the supplied formulation.

Quality and Grades

Item-specific facts (from Product Data)

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

General notes on quality for plasmid CRISPR constructs (literature/general)

• Sequence verification: High-quality sgRNA plasmids are typically sequence-confirmed across the guide region, scaffold, and junctions. Verify with Sanger or NGS before critical experiments.
• Endotoxin levels: Low-endotoxin preparations are preferred for mammalian transfection or in vivo delivery; specifications (EU/mg) vary by supplier. Not specified for this item.
• Topology: Supercoiled fraction affects transfection efficiency. Analytical agarose gel or capillary electrophoresis can assess isoform distribution.
• Residual contaminants: Protein, RNA, salts, and solvent carryover can reduce performance. A260/A280 and A260/A230 ratios give quick assessment; functional testing (transfection titer, editing efficiency) is recommended.
• Stabilizers/buffers: Suppliers may include Tris/EDTA or similar. Specific buffer composition for this item is not specified; refer to CoA/Spec Sheet.
• Traceability: Lot-specific CoA, sequence map, and restriction digest patterns are standard quality documents; consult these for this item’s definitive specifications.

Reaction and Applications

Item-specific facts (from Product Data)

• Product name indicates an sgRNA targeting YTHDF2 within a pLenti backbone; specific cloning sites, promoters, selection markers, and guide sequence are not specified here.

General applications (literature/general)

• Gene editing: When co-expressed with Cas9/Cas12a in mammalian cells, the sgRNA directs site-specific DNA cleavage in YTHDF2, enabling gene knockout, knock-in (with donor template), or functional interrogation via indels.
• Lentiviral delivery: pLenti backbones facilitate stable integration after packaging with helper plasmids (e.g., gag/pol, VSV-G) and transduction of a wide range of cell types. Use BSL-2 practices for viral work.
• Mechanistic studies: YTHDF2 encodes an m6A reader implicated in mRNA decay. Editing this locus supports studies of RNA modification pathways, post-transcriptional regulation, and pathway genetics in cell models.

Practical tips (general)

• Guide validation: Confirm on-target efficiency in your cell line using T7E1, Amplicon-Seq, or ICE/TIDE analysis. Assess potential off-targets in silico and empirically if critical.
• Controls: Include non-targeting and positive-control sgRNAs; rescue with sgRNA-resistant cDNA where appropriate.
• Delivery: Optimize transfection/transduction conditions (cell density, reagent, MOI, antibiotic selection) per cell line. Verify expression of required Cas nuclease if not encoded by the same vector.

Reaction Conditions

Item-specific, tested conditions are not provided for this product.

General guidance for typical workflows (literature/general)

• Transfection (plasmid delivery): Use a DNA:transfection reagent ratio optimized per cell line (e.g., 1–3 µg DNA per well of a 6-well plate). Maintain cells at recommended confluency (typically 50–80%). Replace media after 12–24 h if reagent is cytotoxic.
• Lentiviral packaging: Co-transfect pLenti transfer plasmid with compatible packaging plasmids in HEK293T cells; harvest supernatant at 48–72 h, filter, and use fresh or aliquot and freeze at −80°C. Titrate virus before experimental use. Perform under BSL-2.
• Transduction: Add virus to target cells with polycations (e.g., polybrene) if compatible; select with the encoded antibiotic marker after 48–72 h. Optimize MOI to balance single-copy integration and coverage.
• Editing window: For nuclease-based editing, assess indels 48–96 h post-delivery; for pooled selection, extend as needed. Validate on-target activity and evaluate predicted off-targets when essential to study aims.

Note: Parameters above are general literature practices and not item-specific specifications.

Safety and Handling

Item-specific facts (from Product Data)

• Storage conditions: Store at −80°C.
• Shipping: Dry ice packs + Cold packs.
• GHS classification, signal word, H-statements, pictograms: Not specified for this item; refer to SDS.
• Research use note: For research use only.

General guidance (literature/general)

• Biosafety: Plasmid DNA containing lentiviral elements should be handled in accordance with institutional biosafety guidelines. Work with plasmid DNA itself is typically BSL-1; production and handling of lentiviral particles derived from this plasmid requires BSL-2 (or higher) practices and approvals.
• PPE: laboratory coat, disposable nitrile gloves, and eye protection. Use biological safety cabinet when handling materials that may generate aerosols or when working with viral packaging.
• Incompatibilities: Avoid nucleases (DNase) and contaminated solutions; use nuclease-free, sterile buffers and tips. Avoid strong acids/bases and oxidants that can degrade DNA.
• First aid (overview): If exposure to additives/buffers occurs, rinse skin/eyes with water. If inhalation of aerosolized material occurs, move to fresh air. Seek medical attention per institutional policies. Defer to SDS for authoritative information.
• Decontamination: Surfaces and tools can be decontaminated using freshly prepared 10% bleach followed by ethanol; dispose of DNA-containing waste according to biosafety protocols.

Solvent Selection

This section is not typically applicable to plasmid DNA reagents.

• Item-specific solvent specifications are not provided. Plasmid DNA is handled in aqueous, nuclease-free buffers (e.g., water, TE) rather than organic solvents.
• For formulation/handling guidance, see Storage & Reconstitution and Reaction & Applications.

Storage and Reconstitution

Item-specific facts (from Product Data)

• Storage: Store at −80°C.
• Shipping: Dry ice packs + Cold packs.

General guidance (literature/general)

• Upon receipt: Keep frozen; minimize time at ambient temperature. Document lot and any provided concentration.
• Aliquoting: Thaw once on ice, mix gently, and prepare single-use aliquots in sterile, nuclease-free tubes to avoid repeated freeze–thaw cycles that can shear DNA.
• Buffering: If lyophilized, reconstitute in nuclease-free water or TE buffer to a convenient working concentration (e.g., 0.1–1 µg/µL). If supplied in solution, maintain the provided buffer; do not dilute below concentrations that risk adsorption to plastics.
• Storage durations: Working aliquots at −20°C (short term, weeks) and master stocks at −80°C (long term, months to years). Avoid frost-free freezers and frequent door openings.
• Handling: Use sterile, DNase-free tips and tubes; wipe benches with 10% bleach then ethanol before work.

Any specific buffer composition, concentration, or reconstitution volume for this item is not specified; refer to the CoA/Spec Sheet for definitive instructions.

Structure and Identity

Item-specific facts (from Product Data)

• SKU: P751896
• Product name: pLenti-YTHDF2-sgRNA
• Type: Plasmid/CRISPR guide RNA construct in a lentiviral backbone (inferred from name)
• 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 large nucleic acid constructs.
• SMILES: Not applicable to large nucleic acid 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 description (literature/general)

• pLenti backbones are lentiviral-derived plasmids engineered for cloning and expression; when paired with packaging plasmids they enable production of replication-incompetent lentiviral particles.
• “YTHDF2-sgRNA” denotes a single guide RNA designed to direct Cas nucleases to sequences within the YTHDF2 gene. The plasmid typically contains: a U6 or similar Pol III promoter driving the sgRNA, a gRNA scaffold, and a lentiviral transfer backbone supporting viral packaging elements (e.g., LTRs, Ψ), plus selection markers. Exact features for this item are not specified here.
• 2D structural description: circular double-stranded DNA plasmid comprising standard vector modules (promoter, gRNA cassette, selection marker, origin). No small-molecule structural representation is applicable.

Synthetic Utility

In the context of molecular biology, “synthetic utility” refers to genetic engineering workflows rather than organic synthesis.

General utility (literature/general)

• Modularity: The sgRNA cassette may be swapped or multiplexed to target different loci; cloning strategies often use type IIS restriction enzymes or Gibson/Golden Gate assembly.
• Functional genomics: Enables loss-of-function studies, pooled CRISPR screening (if compatible with library cloning), and epistasis mapping when combined with complementary perturbations.
• Derivatization: The pLenti backbone can be adapted for CRISPRi/a by replacing Cas9 nuclease with dCas9-KRAB or dCas9-activator systems; the same guide sequence can repress or activate gene expression depending on the effector (general guidance).
• Readouts: Downstream validation may include qPCR, RNA-seq, Western blot, and phenotypic assays; on-target edits are quantified by amplicon sequencing or mismatch assays.

Item-specific vector map, cloning sites, and selectable markers are not specified for this product; consult CoA/Spec Sheet for definitive features.

Target Specificity

Item-specific facts (from Product Data)

• Intended target indicated by product name: YTHDF2 gene.
• Guide sequence, PAM context, species, and off-target assessment: Not specified for this item; refer to CoA/Spec Sheet if available.

Notes (general)

• sgRNA efficacy and specificity depend on guide sequence, genomic context, nuclease type, and cell line. Empirical validation is recommended for each application.

Preguntas frecuentes

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-YTHDF2-sgRNA); cross-reactivity in untested species may exist but is not guaranteed. Contact technical support for specific species inquiries.