작성일 : 12-10-28
[AparnaBio] Nanoparticles를 이용한 in vivo transfection 시약
InVivoPlex® / RNAi Nanoparticle System for Murine Cancer Models
InVivoPlex®/Nanoparticles를 이용한 in vivo transfection 시약
RNAi Nanoparticle System for Murine Cancer Models
NanoElectroPlex라고 하는 약물전달 기술을 이용한 in vivo transfection에 시약입니다. InVivo Plex는 self-assembling을 갖는
RNAi nanoparticles로서 혈관투과성, Enhanced Permeability and Retention(EPR) 효과를 이용하여 효율적으로 종양 조직에
핵산을 도입 할 수 있습니다. Enhanced Permeability and Retention(EPR) 효과에 의해 vascularized tumors에 selective
localization 하도록 입자의 크기가 최적화되어 있으며, protective coat 되어 있습니다.
Features ;
  • Simple to use kit that forms siRNA nanoparticles <50 nm, and stable for 1 week
  • Highly selective uptake by vascularized murine tumor models, minimual lung uptake
  • Low toxicity and low levels of cytokine induction, minimizing off-target effects
  • Clinical relevance: multiple dose regimens, dose response, effect simulates targeted therapy
  • Delivers siRNA oligonucleotides, plasmids, and combinations
Tumor Targeted, Therapeutic RNAi Nanoparticle Capabilities ;
  • Tumor Selectivity : Delivery is directed to vascularized tumor tissue, away from organs
  • Repeat Dosing : Administer 3 times per week, without toxicity
  • Dose Response : Evaluate dosages from 1 mg/kg to 6 mg/kg
  • Many Models : Many murine cancer models exhibit EPR active tumors
  • Multiple Cargos : siRNA, plasmids, and combinations can be delivered
Top : InVivoPlex® luciferase plasmid nanoparticles given iv selectively target vascularized subcutaneous tumors in
xenograft murine cancer models, without complicating lung or other organ activity typical of non-targeted in vivo
transfection reagents.
Bottom : InVivoPlex® siRNA nanoparticle treatment by iv, 3 times per week for two weeks, inhibits xenograft tumor
growth. Left: pan human siRNA (Qiagen). Right: InVivoActTM hVEGF - equivalent to inhibition by the approved hVEGF
mAb therapeutic.
Applications ;
  • Study in vivo function of tumor genes in a clinically relevant fashion
  • Prioritize the most promising gene targets for drug discovery
  • Study multi-gene inhibition or gene interactions with chemotherapy
  • Validate candidate RNAi or gene cancer therapeutics in vivo
Simple to Use , Complete Kit ;
  • Simple Assembly : Add aqueous nucleic acid to NP Reagent Vial drop wise while stirring
  • Cargo Flexibility : You select the cargo: siRNA, shRNA, or expression plasmid
  • Convenient : Nanoparticles can be prepared once each week for administration
  • Complete Kit : Mixing system and all buffers, including Vehicle Control, are provided
  • Accessory Kits : Specialized RNAi to facilitate tumor uptake and gene activity studies
※ 분석 전에 종양 형성 마우스의 EPR 활성을 측정하거나 또는 EPR 활성이 증가하도록 조절하십시오. EPR 활성의 평가는
   InVivo Trak을 사용하거나, InVivo Plex와 reporter gene plasmid를 함께 사용하는 것이 좋습니다. (InVivo Track에 대한
   제품 소개가 바로 될 예정입니다.)
Points To Consider
Determine murine tumor model EPR activity :
Studies are recommended to first evaluate EPR activity of tumor models, or to adjust tumor model parameters to
increase EPR activity. This can be accomplished using InVivoTrak™, fluorescent dye labeled agents designed for tumor
uptake studies, or InVivoPlex® with a reporter gene plasmid. Initial studies with these agents can address concerns of
variability of cancer models among labs, and variation related to tumor vasculature that influence activity of the
nanoparticle targeting the tumor tissue. The parameters of the protocol can be established for efficacy studies,
confirming that InVivoPlex® nanoparticles actually target oligonucleotide delivery selectively to the tumor tissue.
Use InVivoPlex® to evaluate gene or protein inhibition in vivo :
Use of InVivoPlex® to evaluate gene and protein inhibition will likely require repeated treatments, and is expected to
give 20-40% inhibition of targeted mRNA. We recommend three doses, e.g. Monday, Wednesday, and Friday, assayed
the following Monday. Note, even modest inhibition of key proteins can elicit a significant effect - see the figure on page 2,
showing equivalent inhibition of an NSCLC xenograft tumor model by Avastin® or InVivoPlex® delivery of InVivoActTM-
hVEGF. The genes and proteins showing the greatest effect from low levels of inhibition are likely to represent the
most potent points for therapeutic intervention.
Evaluate experimental therapeutic targets in vivo with InVivoPlex® :
Use of InVivoPlex® to evaluate candidate therapeutic targets identified in vitro with RNAi agents will usually require
repeated doses to show an effect on tumor behavior, and typically requires multiple treatment groups with negative
controls  and ideally a positive control RNAi agent. InVivoActTM control RNAi have been developed to provide negative
and positive controls, as shown in results on page 2 where InVivoPlex® with InVivoActTM-hVEGF inhibited H460 tumor
growth as well as the approved mAb therapeutic while InVivoActTM-Neg had no effect compared to buffer control
treatment. These agents can be used to facilitate validation of candidate targets with even greater potency in similar
tumor cell pathways.
Use InVivoPlex® to evaluate gene combinations or interaction with chemotherapeutics :
The ratio of siRNA or plasmid combinations for multi-target studies will depend on their relative potency, e.g. plasmid
expression or siRNA gene inhibition activity. Combination studies can be initiated with equal amounts in studies
characterizing activity, permitting adjustment of the ratio based on relative potency. When combining plasmid gene
expression with siRNA gene inhibition, we found that about one third siRNA by weight provided both gene
expression and plasmid gene inhibition by an inhibitory siRNA, after 24 hours. To study synergy or interference of target
genes with chemotherapy, multidose regimen studies can be performed by including groups with separate and
combined siRNA and chemotherapy treatments.
Regulatory issues for performing InVivoPlex® studies :
Use of animals for scientific research usually is subject to national and in some cases regional or local laws. For
laboratories in the US AparnaBio can help provide draft protocol language and guidance as well as links to information
available from the US Government, and gladly discuss how use of InVivoPlex® can be described to review committees,
including a description of how its use is consistent with established humane use of experimental animals.
Product Summary
InVivoPlex® Kits enable translation of RNAi studies to murine models of cancer, with the objective to allow studies
producing clinically relevant information on tumor gene function, candidate targets for therapeutic intervention, and in
vivo validation of RNAi therapeutic candidates. The InVivoPlex® Kits supply all the required buffers and the nanoparticle
forming reagent, along with everything needed for specialized mixing with user supplied RNAi agents forming EPR
targeted nanoparticles that localize in vascularized tumors in murine models of cancer, as well as vehicle control buffer for a control treatment group of mice. An important advantage of the reagent is the feature that it allows the user to
prepare the nanoparticles with their experimental RNAi agent, and the capability to use a wide variety of RNAi agents as
well as to perform combination RNAi studies. InVivoPlex® Kits are available for preparation of nanoparticle samples for
treatment of small, medium, or large groups of mice. Kits for other group sizes can be provided upon request.
To use InVivoPlex®, we recommend starting with a tumor uptake study to insure EPR activity of the specific murine
cancer model to be used, or its optimization for EPR activity. The InVivoTrakTM fluorescent labeled companion products
are available to facilitate these studies. The InVivoTrakTM is available with either red or near IR fluorescent labels, and
can be used with tumor homogenates in a fluorescence plate reader or with an NIR in vivo imaging instrument. InVivo
TrakTM fluorescent tracer products are designed for use with small size group studies. The InVivoTrakTM Oligo products
are designed for use with the small InVivoPlex® kit, to make fluorescent labeled nanoparticles for testing EPR tumor
uptake. InVivoTrakTM is available with either red or near IR fluorescent labels. The InVivoTrakTM Carrier products are
designed for use with unlabeled RNAi or with InVivoTrakTM oligo products for dual label tracer studies.
If studies of tumor gene inhibition with InVivoPlex® are desired, we recommend a one week, three dose protocol.
For studies of RNAi activity on tumor growth or other phenotype, we recommend multi-dose studies over several weeks with multiple treatment groups that include negative and positive controls. The InVivoActTM negative and positive RNAi agent
companion products are available to facilitate these studies. InVivoActTM products are designed for use with the
large InVivoPlex® Kit, and are available in forms to provide a negative RNAi treatment and to provide positive RNAi
treatments that inhibit a clinically validated therapeutic target gene, such as VEGF or EGFR.
Ordering informations ;
  1. IVP-T-S   InVivoPlex-Tumor RNAi Nanoparticle Kit, Small(마우스 6~7마리의 양)  
  2. IVP-T-M  InVivoPlex-Tumor RNAi Nanoparticle Kit, Medium(마우스 8~9마리의 양)
  3. IVP-T-L   InVivoPlex-Tumor RNAi Nanoparticle Kit, Large(마우스 10~12마리의 양)    
※ 본 제품은 연구용입니다. 임상 용도로는 사용할 수 없습니다.
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