hsa-miR-4643 mimic

Cat. No.: H1459905

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Übersicht

hsa-miR-4643 mimics are small, chemically synthesized double-stranded RNAs that mimic endogenous miRNAs and enable miRNA functional analysis by up-regulation of miRNA activity.

Specifications

Storage

Store at -20°C

Verschickt in

Ice chest + Ice pads

Dieses Produkt erfordert Kühlkettenversand. Grundversand und andere Economy-Optionen sind nicht verfügbar.

Product Line

Biologische Produkte

Application Protocols

The following protocols are general best‑practice guidance for miRNA mimics; they are not item‑specific. Parameters should be optimized for each cell type and transfection reagent.

A. Rapid transfection workflow (adherent cells)

1. Plate cells to 30–60% confluency on transfection day.
2. Prepare complexes: Dilute mimic in serum‑free medium; in a separate tube, dilute transfection reagent. Combine, gently mix, and incubate 10–15 min.
3. Add complexes dropwise to cells (target final 5–50 nM mimic). Swirl gently.
4. Incubate 4–6 h; optionally replace with fresh complete medium.
5. Harvest at 24–72 h for RT‑qPCR, luciferase, or protein assays.

B. Dual‑luciferase target validation

• Co‑transfect reporter plasmid containing candidate 3′UTR (WT) and a control plasmid. In parallel, transfect a seed‑mutant 3′UTR.
• Read firefly/Renilla activities 24–48 h post‑transfection; calculate normalized repression relative to non‑targeting mimic.

C. RT‑qPCR assay for target repression

• Isolate total RNA with miRNA‑compatible kits; perform reverse transcription with random hexamers/oligo‑dT for mRNA targets.
• Quantify target mRNA and housekeeping controls; analyze fold change (ΔΔCt).

Controls and QA

• Include non‑targeting mimic, positive control miRNA, and mock transfection. Monitor cytotoxicity (e.g., resazurin).

Note: Item‑specific dilutions, buffer composition, and validated cell lines are Not specified for this item; refer to CoA/Spec Sheet.

Biological Roles

General background (microRNAs)

• microRNAs (miRNAs) are ~22‑nt noncoding RNAs that guide Argonaute‑containing RISC to partially complementary sites in target mRNAs, leading to translational repression and/or mRNA decay. Target recognition is driven primarily by the seed sequence (positions 2–8 of the guide strand).

hsa‑miR‑4643 context

• Specific biological roles and validated targets of hsa‑miR‑4643 are an active area of study in the literature. Without the exact sequence and curated target list for this product, definitive target/pathway annotations are not provided here.

Research use of mimics

• A miR‑4643 mimic supplies the mature guide strand to cells, bypassing endogenous biogenesis steps (Drosha/Dicer) and enabling controlled gain‑of‑function. This facilitates discovery of direct targets (via reporter assays and CLIP‑based approaches) and indirect effects (via transcriptomics/proteomics).

Best practices

• Integrate computational predictions (multiple algorithms) with experimental validation (reporter assays, RT‑qPCR, western blot). Use seed‑mutant controls and rescue experiments to distinguish on‑target from off‑target effects.

Note

• No clinical/diagnostic claims are made. This product is for research use only. Item‑specific sequence, modifications, and any known targets are Not specified for this item; refer to CoA/Spec Sheet and current literature.

Buffer Applications

Relevance: Although not a buffer component, this oligonucleotide requires appropriate buffers for dissolution, storage, and use.

Common buffers (general guidance)

• Reconstitution: RNase‑free water or 1× TE (10 mM Tris‑HCl, 1 mM EDTA, pH 7.5–8.0). TE can enhance stability during frozen storage by chelating divalent cations.
• Working solutions: RNase‑free water or serum‑free transfection medium for complex formation with lipid/polymer reagents. After complexation, dilute into complete growth medium.
• Avoid Mg2+/Mn2+ in long‑term stocks, which can catalyze RNA hydrolysis.

Preparation tips

• Warm gently to room temperature; briefly spin to collect material; mix by gentle pipetting—avoid vigorous vortexing that can shear duplexes.
• Use low‑binding, RNase‑free tubes and tips. Prepare small aliquots to minimize freeze–thaw.

pH considerations

• Neutral to slightly basic pH (7.0–8.0) is generally preferred for stability and compatibility with transfection reagents.

Item-specific formulations

• Any proprietary buffer system or stabilizers are Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Applicability: Green chemistry considerations are limited for nucleic acid mimics, which are used at low mass and in aqueous systems. Nevertheless, choices around workflows and consumables can reduce environmental impact.

Strategies (general guidance)

• Prefer aqueous systems: Prepare and use stocks in water/TE; avoid organic cosolvents unless essential for delivery.
• Low‑toxicity delivery: Select transfection reagents with lower cytotoxicity and reduced solvent content; optimize to minimize reagent amounts.
• Waste minimization: Plan experiments to right‑size reaction volumes; pool biological replicates when statistically appropriate.
• Reusables: Employ reusable cold blocks and racks; minimize single‑use plastics where contamination risk is manageable.
• Energy efficiency: Consolidate freezer storage (-20 °C) space and use insulated transport to reduce dry ice consumption for short transfers.

Comparison (conceptual)

• Synthetic mimics vs plasmid overexpression: Synthetic mimics avoid bacterial culture, antibiotic use, and DNA purification reagents, reducing certain waste streams; however, they entail solid‑phase synthesis waste upstream. Plasmids enable renewable supply but require additional cell culture resources.

Item-specific greener options

• Not specified for this item; select methods consistent with your lab’s green chemistry policies and validation data.

Pharmaceutical Uses

This product is supplied strictly for research use only and is not intended for human or veterinary use, diagnostic procedures, or as a pharmaceutical ingredient.

Context (general information)

• Synthetic oligonucleotides can serve as active substances or excipients in regulated settings; however, such applications require GMP manufacturing, extensive characterization (identity, purity, residual solvents, endotoxin), and regulatory filings. None of these are specified or implied for this item.

Formulation relevance in research

• For in vitro studies, miRNA mimics are typically formulated with transfection reagents into lipid or polymer complexes immediately before use. No pharmacopeial status is claimed.

Item-specific regulatory/quality status

• Not specified for this item; refer to CoA/Spec Sheet if any compliance statements are provided.

Physical Properties

Item-specific values

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• pKa, logP, refractive index: Not applicable/Not typically defined for oligonucleotides; refer to CoA/Spec Sheet if provided.

General physical characteristics of short RNA oligonucleotides (literature/guidance)

• State: Solid when lyophilized; readily soluble in nuclease‑free water or TE buffer owing to the polyanionic phosphate backbone.
• Solubility: Freely soluble in aqueous buffers (e.g., water, 1× TE, 1× PBS without Mg2+ for stock preparation). Poorly soluble in organic solvents.
• UV absorbance: Strong absorption at 260 nm; concentration often determined by A260 using sequence‑specific extinction coefficients.
• Thermal behavior: Duplex melting temperature (Tm) depends on sequence, length, salt, and any modifications; not specified for this item.
• Stability: Sensitive to hydrolysis and ribonucleases; stability improved at low temperature, neutral to slightly basic pH, and in RNase‑free conditions.

Handling implications

• Prepare and store stocks in RNase‑free solutions. Avoid repeated freeze–thaw; aliquot as single‑use portions.

Caution

• Where exact numerical specifications (e.g., extinction coefficient, Tm, residual moisture) are required, Not specified for this item; refer to CoA/Spec Sheet.

Quality and Grades

Item-specific grade/purity

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

Context for oligonucleotide quality (general guidance)

• Common purities: Desalted (crude), cartridge‑purified, HPLC‑purified, or PAGE‑purified. Higher purities reduce truncated sequences and are preferred for functional assays.
• Identity/assay: Typical QC may include MALDI‑TOF or ESI‑MS for mass confirmation and analytical HPLC for purity (literature). Some vendors report OD260 and coupling yields. These are not specified for this item unless stated on the CoA.
• Modifications: miRNA mimics may carry terminal or backbone modifications (e.g., 2′‑OMe, 2′‑F, phosphorothioate, duplex asymmetry) to enhance stability and activity. Any such details are Not specified for this item.
• Endotoxin/RNase: For cell‑based work, low endotoxin and RNase‑free handling are critical; verify on the CoA if required by your application.

Practical selection tips

• Choose purification grade commensurate with assay sensitivity (e.g., HPLC for reporter assays and transcriptomics; desalt may suffice for screening). Confirm sequence and modifications on the supplied documentation.

Reaction and Applications

Scope: As a functional genomics reagent, the hsa‑miR‑4643 mimic is used to transiently increase miR‑4643 activity in cells, enabling pathway interrogation and target validation. It is not a small‑molecule reagent for chemical reactions.

Representative research applications (general, literature)

• Gain‑of‑function studies: Ectopic elevation of the guide strand to assess phenotypic effects and downstream transcript changes.
• Target validation: Co‑transfection with 3′UTR luciferase reporters bearing wild‑type vs seed‑mutated sites to confirm direct targeting.
• Pathway mapping: RNA‑seq or microarrays to profile differential gene expression upon mimic transfection; enrichment analyses for affected pathways.
• High‑content/CR screens: Use in arrayed screens to probe gene networks when combined with imaging or viability readouts.
• Rescue/epistasis experiments: Combine with siRNA/CRISPR perturbations to dissect regulatory hierarchies.

Practical considerations

• Dosing: Typical miRNA mimic working concentrations are 5–50 nM; optimize per cell line and endpoint (general guidance).
• Timing: Phenotypes often manifest 24–72 h post‑transfection; mRNA changes may appear earlier than protein changes.
• Controls: Use non‑targeting mimic controls, positive controls (well‑characterized miRNAs), and seed‑mutant reporters for specificity.
• Readouts: RT‑qPCR for primary targets, western blot for protein, reporter assays, and phenotypic assays.

Note

• Specific performance characteristics for this item are Not specified; consult the CoA/Spec Sheet and optimize empirically.

Reaction Conditions

Chemical reaction conditions are not applicable. The relevant “conditions” pertain to cell transfection and assay timing. The following are general literature‑based guidance for miRNA mimics; optimize empirically for your system.

General transfection parameters (guidance)

• Cell lines: Adherent lines (e.g., HEK293, HeLa) and many suspension lines can be transfected; choose a delivery reagent validated for miRNA/siRNA.
• Dose: Start with 5, 10, 25, and 50 nM final mimic concentration; select the lowest dose that achieves a robust on‑target response.
• Complexation: Mix mimic with transfection reagent in serum‑free medium; incubate 5–20 min to form complexes; add to cells and return to complete medium after 4–6 h if required.
• Incubation: 24–72 h for mRNA/protein readouts; up to 96 h for phenotypes. Monitor cell viability.
• Controls: Non‑targeting mimic, positive control miRNA, and mock transfection. Include seed‑mutant 3′UTR reporters for specificity.

Assay readouts

• RT‑qPCR for predicted targets and pathway markers; dual‑luciferase reporter assays for direct binding; western blot or flow cytometry for protein changes; RNA‑seq for global effects.

Note

• No item‑specific, validated conditions are provided here. Not specified for this item; refer to CoA/Spec Sheet and optimize conditions per cell type and reagent.

Safety and Handling

GHS and hazard data

• Signal word: Not specified for this item; refer to SDS.
• H‑statements: Not specified for this item; refer to SDS.
• GHS classification/pictograms: Not specified for this item; refer to SDS.

General safety guidance for RNA oligonucleotides (literature/practice)

• Expected hazards: Non‑volatile, non‑combustible polyanions; typically low acute toxicity. Main risks are from dust generation (if lyophilized) and contamination with RNases compromising experiments.
• Personal protective equipment: Laboratory coat, safety glasses, and disposable nitrile gloves. Work in a clean area dedicated to RNA handling; change gloves frequently to minimize RNase contamination.
• Engineering controls: Use RNase‑free consumables; consider a clean bench or PCR workstation for critical steps.
• Incompatibilities: Avoid strong acids/alkalis that promote backbone cleavage; avoid ribonucleases (ubiquitous on skin and common labware). Keep away from nucleases, metal ion contaminants, and microbial contamination.
• First aid (general): If contacted with eyes/skin, rinse with water. If inhaled as dust, move to fresh air. If ingested, rinse mouth with water. Seek medical attention as appropriate and present SDS.
• Spill/cleanup: Wipe with aqueous detergent solution; dispose according to institutional guidelines for nonhazardous biological chemicals.

Authoritative source

• Always defer to the product‑specific Safety Data Sheet (SDS) for definitive hazard, handling, and disposal instructions.

Solvent Selection

Applicability: As an RNA oligonucleotide, this product is not used as a reaction solvent. Solvent selection pertains to dissolution and handling for biological assays.

Recommended media (general guidance)

• Primary: Nuclease‑free water (RNase/DNase‑free). Prepare concentrated stocks (e.g., 10–100 µM) for aliquoting.
• Stabilizing buffers: 1× TE (10 mM Tris‑HCl, 1 mM EDTA, pH 7.5–8.0) can improve long‑term stability by chelating divalent metals that catalyze hydrolysis.
• Avoid: Strongly acidic/basic solutions; repeated freeze–thaw in dilute solutions; solutions containing RNases or Mg2+ during storage.

Miscibility/compatibility (general)

• Aqueous systems: Fully miscible. Compatible with most cell culture media upon dilution; verify with your transfection reagent.
• Organic solvents: Generally insoluble or unstable; avoid DMSO unless validated for your protocol and compatible with transfection chemistry.

Comparison (use‑case oriented)

• Water vs TE: Water is suitable for short‑term and when EDTA is undesirable (e.g., Mg2+‑dependent enzymes later). TE is preferred for freezer stocks to mitigate metal‑catalyzed degradation.

Note

• Any item‑specific solvent constraints are Not specified for this item; refer to CoA/Spec Sheet.

Storage and Reconstitution

Item-specific instructions

• Storage conditions: Store at -20°C (per product data). Protect from moisture and RNase contamination.
• Shipping: Shipped in an ice chest with ice pads (per product data). Inspect upon arrival; if thawed, promptly return to recommended storage after verifying integrity.

General reconstitution guidance for miRNA mimics

• Reconstitution solvent: RNase‑free water or 1× TE buffer, pH 7.5–8.0.
• Stock concentration: Commonly 10–100 µM; choose based on anticipated working range and aliquot size.
• Technique: Briefly centrifuge the vial to collect material; add calculated volume; gently pipette to dissolve; avoid vigorous vortexing. Allow 10–15 min at room temperature to fully solubilize.
• Aliquoting: Prepare single‑use aliquots in low‑binding, RNase‑free tubes to avoid repeated freeze–thaw cycles.

Storage after reconstitution

• Short term (≤1 week): 2–8 °C.
• Long term: -20 °C or -80 °C for months; TE stocks often show improved stability vs water. Avoid frost‑free freezers that cycle temperatures.

Stability notes

• Protect from nucleases; use RNase‑free consumables and change gloves frequently.
• Do not store working dilutions for extended periods; prepare fresh as needed.

Any item‑specific concentration, stabilizers, or shelf‑life claims are Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

Overview: hsa-miR-4643 mimic is a synthetic, double‑stranded RNA oligonucleotide designed to functionally mimic the endogenous human microRNA miR‑4643 in cultured cells and other research systems.

Item-specific identifiers

• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES: Not applicable to large, sequence-defined oligonucleotides.
• InChIKey: Not applicable to large, sequence-defined oligonucleotides.
• CAS / CID: Not specified for this item; refer to CoA/Spec Sheet.

Structural features (general, literature)

• Class: microRNA mimic (typically a short RNA duplex, ~21–23 nt) with a guide strand matching the mature hsa‑miR‑4643 sequence and a passenger strand designed for RISC loading efficiency.
• Functional groups: Repeating ribose–phosphate backbone with nucleobases (A, U, G, C). May include standard 3′/5′ termini; any chemical modifications are not specified for this item.
• Stereochemistry: Natural ribose stereochemistry at each chiral center; polyanionic phosphate backbone under physiological pH.

2D structural description (general)

• Linear polynucleotide chains (two strands) forming a short A‑form RNA duplex with characteristic base‑pairing (Watson–Crick) and 5′/3′ termini. The guide strand’s seed region (nt 2–8) drives target recognition in RISC.

Notes

• Exact sequence, length, and any stabilization chemistries are Not specified for this item; refer to CoA/Spec Sheet.
• For sequence‑level work (e.g., target prediction), consult the specific sequence supplied on the product documentation.

Synthetic Utility

Not typically applicable. This product is a biological oligonucleotide reagent used for gene regulation studies rather than as a building block or reagent in chemical synthesis.

For researchers seeking nucleic acid chemistry details (general)

• Oligonucleotide synthesis employs solid‑phase phosphoramidite chemistry; however, the product is supplied post‑synthesis for biological use. Any monomer composition or modification chemistry for this specific item is Not specified; refer to CoA/Spec Sheet if available.

Target Specificity

Item-specific targeting details

• Designed intent: To mimic the endogenous human miRNA hsa‑miR‑4643. Exact sequence, seed region, and any chemical modifications are Not specified for this item; refer to CoA/Spec Sheet.

General specificity principles for miRNA mimics (literature)

• Primary determinant: The seed region (nt 2–8 of the guide strand) directs binding to complementary sites in target 3′UTRs, 5′UTRs, or CDS.
• Off‑target potential: Seed‑mediated off‑targets can occur in transcripts with partial complementarity; passenger‑strand loading may also cause unintended effects if not suppressed by design.
• Context: Target site accessibility, flanking nucleotide composition, and multiple site occurrences modulate repression.

Recommended validation

• Reporter assays with wild‑type vs seed‑mutated sites.
• RT‑qPCR/western blot for candidate targets.
• Rescue experiments: Overexpress target ORF lacking the 3′UTR site to confirm specificity of observed phenotypes.

Note

• No antigen/epitope/clone concepts apply; this is not an antibody. Species reactivity is expected to be human‑centric given “hsa” designation, but functional cross‑reactivity depends on sequence conservation; consult current literature.

Häufig gestellte Fragen

How should this product be stored?

Store at ?20 °C. Freezer storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

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.