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友善列印TransIT®-Lenti Transfection Reagent

專為提升慢病毒 (lentivirus) 產量而設計的細胞轉染試劑。產品特點包含:

  • 生產系統彈性:適用於懸浮型與貼附型 HEK293 細胞生產 lentivirus。
  • 高病毒產率:具有優於 Lipofectamine® 2000、Lipofectamine® 3000、25kDa PEI、CaPO4 的 lentivirus 生產效率。
  • 操作簡易:有別於其他市售品牌與傳統轉染試劑, TransIT®-Lenti 無須更換培養基,並且只需要收集一次病毒液即可獲得高病毒力價。
  • 高安全性:成份不含任何動物來源物質,大幅降低細胞污染的可能性。

 

High Functional Titers with TransIT®-Lenti Transfection Reagent. Adherent 293T/17 cells were transfected in a 6-well plate with pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION® (1:1 ratio, 2 µg/well) with the following reagents: TransIT®-Lenti (3:1, vol:wt), Lipofectamine® 2000 (3:1), Lipofectamine® 3000 (3:1:1), 25 kDa PEI (6:1), or CaPO4 precipitation (4 µg pDNA/well). The supernatant was harvested, filtered (0.45 µm), and titered using 293T/17 cells. Lentivirus transductions were performed in the presence of 8 µg/ml TransduceIT™ and GFP expression was measured 72 hours post-transduction using guava easyCyte™ 5HT Flow Cytometer. Error bars represent triplicate transfection complexes titered individually. Functional titers were calculated using virus dilutions with less than 20% GFP positive cells.

High Transduction Efficiency with Unconcentrated Lentivirus Using TransIT®-Lenti. (A) Lentivirus was produced with the TransIT®-Lenti Transfection Reagent (3:1, vol:wt) or Lipofectamine® 2000 using pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION®). The supernatant was harvested, filtered (0.45 µm), and frozen. Lentivirus transductions were performed 5 days post-plating with iCell® Motor Neurons (Cellular Dynamics International). For bothTransIT-Lenti and Lipofectamine® 2000, one microliter of unconcentrated supernatant was added per well of a 96-well plate. GFP efficiency was measured 72 hours post-transduction using guava easyCyte™ 5HT Flow Cytometer. Error bars represent the SEM of duplicate wells. (B) iCell® Motor Neurons were plated in Ibidy 35mm dishes and transduced with lentivirus produced using the TransIT®-Lenti Transfection Reagent and MISSION® vectors. Images were captured at 72 hours post-transduction with a Zeiss Axiovert S100 inverted fluorescence microscope using a 63X objective under oil.

Comparison of CaPO4, Lipofectamine® 2000 or TransIT®-Lenti Generated Lentivirus. HIV CMVeGFP Virus was produced in HEK 293FT cells using either CaPO4, Lipofectamine® 2000 or TransIT®-Lenti Transfection Reagent per the manufacturer's protocol. Lentivirus was collected 48 hours post-transfection and concentrated by prolonged centrifugation at 9,000 x g. HT1080cells were infected with a 1:100 or 1:1000 dilution of each concentrated lentivirus. Images (above) were captured 48 hours post-transduction.

High Efficiency Transfection with TransIT®-Lenti Transfection Reagent. Adherent 293T/17 cells were transfected in a 6-well plate format using the pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION® using the TransIT®-Lenti Transfection Reagent (3:1, vol:wt). GFP efficiency was measured at 48 hours post-transfection using guava easyCyte™ 5HT Flow Cytometer. Error bars represent five transfection complexes. Images were captured at 48 hours post-transfection (10X objective) using a Zeiss Axiovert S100 inverted fluorescence microscope. The observed cell rounding and cell-cell fusion is due to high expression of the vesicular stomatitis virus G protein (VSV-G) for pseudotyping the recombinant lentivirus.

Lentivirus Production is Scalable. Adherent 293T/17 cells were transfected in a 12-well, 6-well or 100 mm plate format using the pLKO.1-puro-CMV-TurboGFP™ transfer vector and the Lentivirus Packaging Mix Powered by MISSION® at a 1:1 ratio, and the TransIT®-Lenti Transfection Reagent (3:1, vol:wt). The supernatant was harvested, filtered (0.45 µm), and titered using 293T/17 cells. Lentivirus transductions were performed in the presence of 8 µg/ml TransduceIT™ and GFP expression was measured 72 hours post-transduction using guava easyCyte™ 5HT Flow Cytometer. Error bars represent triplicate transfection complexes titered individually. Functional titers were calculated using virus dilutions with less than 20% GFP positive cells.

 

訂購資訊

Product Pack Size Cat. No.
TransIT®-Lenti Transfection Reagent 0.3 ml MR-MIR6603
0.75 ml MR-MIR6604
1.5 ml MR-MIR6600
5 x 1.5 ml MR-MIR6605
10 x 1.5 ml MR-MIR6606

 

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