MSCVレトロウイルスパッケージング

MSCV retroviral vectors serve as efficient viral vector tools for achieving robust expression of genes of interest in embryonic stem (ES) cells, embryonal carcinoma (EC) cells and hematopoietic stem (HS) cells, along with several other mammalian cell lines. The presence of a strategically designed 5’LTR derived from the murine stem cell PCMV virus in the MSCV retroviral vector contributes to the transcriptional activation of target genes in pluripotent cell lines. This helps to overcome the restricted expression of target genes in ES and EC cells, typically observed with MMLV retroviral vectors.
VectorBuilder has developed a series of proprietary technologies and reagents that have greatly improved MSCV packaging protocols in terms of titer, purity, viability and consistency, especially for the MSCV vector systems used in our vector cloning services. As a result, we have a growing base of highly satisfied customers who come back to us time after time for their MSCV vector cloning and packaging needs.
Click here to read detailed introduction of our MSCV vector systems
Click here to view citations of our MSCV
Click here to view user testimonials of our MSCV
How do I obtain MSCV packaging services?

Click here to view an example of a finished MSCV vector design
Click here to view an example of a MSCV service proposal
Click here to view instructions on designing a vector on VectorBuilder
Click here to view instructions on placing an order on VectorBuilder
Types of MSCV retrovirus offered

We offer packaging services for VSV-G pseudotyped MSCV retrovirus.

価格と出荷までの作業日数

Due to our technological advantage, our MSCV packaging services have the highest quality in the industry and are also faster and more affordable than other suppliers. Price and turnaround time for different scales of MSCV packaging are shown below:

サイズ 推奨の使用系 タイター*と仕様 価格 (税抜き) 出荷日数**
パイロット 培養細胞 >107 TU/ml, 10x 25 ul, HBSS buffer 64,000円 7-14 営業日
中容量 培養細胞 >107 TU/ml, 10x 100 ul, HBSS buffer 89,000円 7-14 営業日
大容量 培養細胞 >108 TU/ml, 10x 100 ul, HBSS buffer 139,000円 7-14 営業日
超純粋大容量

培養細胞とin vivo

>108 TU/ml, 10x 100 ul, HBSS buffer 201,500円 7-14 営業日

* Our titer guarantee only applies to vectors for which the region being packaged into virus (from 5’ LTR to 3’ LTR) is below MSCV cargo limit (8 kb). For sizes over this range, it may still be possible to package the vector into virus, but the titer may be reduced. Additionally, we are not able to guarantee titer for the following vectors:

  • vectors containing sequences that could adversely affect the packaging process such as toxic genes (e.g. proapoptotic genes), genes that compromise the integrity of packaging cells or virus (e.g. membrane proteins that cause cell aggregation), and sequences prone to rearrangements or secondary structures;
  • customer-supplied vectors as we have no control over their quality.

** Estimated turnaround time is the time from production initiation to completion. It does not include waiting time for any customer-supplied materials (e.g. template DNA), QC of such materials, and transit time for shipping final deliverables to the customer.

Deliverables

If you choose pilot, medium or large scale MSCV packaging, each order includes the following deliverables:

  • Aliquots of custom MSCV at selected scale
  • Free: aliquots of control virus*
  • Free: Polybrene (5 mg/ml, 200 ul)

If you choose ultra-purified large scale MSCV packaging (in vivo grade), each order includes the following deliverables:

  • Aliquots of custom MSCV at selected scale
  • "Add-on" purchase (optional): aliquots of ultra-purified control virus*
  • Free: Polybrene (5 mg/ml, 200 ul)
* The control virus is designed to match the biological application of the custom virus and to be used for testing transduction. For example, if the custom virus overexpresses a gene, then the control virus provided will overexpress EGFP. Detailed information of the control virus is shown below:
ベクターシステム コントロールベクター名称 コントロールベクターID カスタムウイルスのパッケージングサイズ コントロールウイルスの仕様 価格(税抜き)
MSCV 遺伝子発現システム pMSCV[Exp]-EGFP:T2A:Puro VB190529-1011paj パイロット >107 TU/ml, 100 ul in HBSS 無料
中容量 >107 TU/ml, 2x 100 ul in HBSS 無料
大容量 >107 TU/ml, 2x 100 ul in HBSS 無料
超純粋大容量 >108 TU/ml, 10x 100 ul in HBSS 76,500円
ウイルス粒子作製、精製、タイター測定

For our MSCV packaging, the transfer vector carrying the gene of interest (GOI) is co-transfected with our proprietary envelop vector encoding VSV-G and packaging vector encoding Gag and Pol. After a short incubation period, the supernatant is collected and centrifuged to remove cell debris and then filtered. Viral particles are subsequently concentrated with PEG. For ultra-purified (in vivo grade) MSCV, viral particles are further purified and concentrated by sucrose cushion centrifugation. We use a qPCR-based approach to measure the functional titer of MSCV.

品質管理

For each MSCV produced by VectorBuilder, the quality control includes functional titer measurement, sterility testing for bacteria and fungi, and mycoplasma detection. If the transfer vector encodes a fluorescent protein, we would transduce cells with the virus and detect corresponding fluorescence. Additionally, for ultra-purified MSCV, we routinely sample virus quality using the endotoxin assay.

ユーザーご提供のトランスファーベクターからのウイルス粒子受託作成について

ユーザーお手持ちのウイルスベクターからウイルス粒子の受託作製も受注しています。 ウイルスベクターはマテリアルサブミッションガイド.プラスミドベクターは、QCチェックとして、マキシプレップ、プラスミドの制限酵素サイトの確認と、サンガーシークエンシングでのシークエンスの確認を行います。一律12500円のQCチェック費用をお見積りに加算させていただいています。 QCチェックに合格後、ご注文のウイルスパッケージングサービスを行います。ウイルスパッケージングのお見積りの作業日数には QCチェックにかかる日数(通常5営業日)は含まれていません。あらかじめご了承ください。

ご質問

よくあるご質問は Vector FAQ からご覧になれます。弊社からご提供させていただいている受託サービスについて、もっと詳細な情報が必要な場合は  ベクタービルダー・ジャパン サービスサポートまでお問合せください。

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