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TransIT
¢ç-AAViator Transfection System (Code MIR 73750)ÀÇ
´ë¿ë·® Á¦Ç° (15L cell culture¿ë)Àº ´ÙÄ«¶óÄÚ¸®¾Æ °í°´Áö¿ø (02-2081-2510,
support@takara.co.kr)·Î ¹®ÀÇ ¹Ù¶ø´Ï´Ù.
- ³ôÀº °¨¿°È°¼ºÀ» °¡Áö´Â °íÇ°ÁúÀÇ AAV »ý»ê¿¡ ÃÖÀû
- HEK293 ¼¼Æ÷ ¹è¾ç ½Ã½ºÅÛ¿¡ ÃÖÀû (adherent ¹× suspension ¹è¾ç ¸ðµÎ Àû¿ë)
- ¿¬±¸¿ë ½Ã¾àºÎÅÍ ÀüÀÓ»ó ´Ü°è, ÀÓ»ó¿ë GMP grade±îÁö ¶óÀξ÷
Á¦Ç° ¼³¸í
TransIT¢ç- AAViator Transfection SystemÀº MirusÀÇ ÀÔÁõµÈ polymer, lipid technologyÀ» ÀÌ¿ëÇÏ¿© °³¹ßµÈ AAV »ý»êÀ» À§ÇÑ ¸ÂÃ㠽ýºÅÛÀÌ´Ù. º» Á¦Ç°À» »ç¿ëÇϸé, Ÿ»çÀÇ Polymer ´ÜÀÏ ½Ã¾à¿¡ ºñÇØ AAV »ý»ê·®À» Áõ°¡½Ãų ¼ö ÀÖ°í, ½±°Ô scale up ÇÒ ¼ö ÀÖÀ¸¸ç, Çö󽺹̵å DNA »ç¿ë·®À» ÃÖ´ë 50% ¾Æ³¥ ¼ö ÀÖ´Â ÀåÁ¡ÀÌ ÀÖ´Ù.
TransIT¢ç- AAViator Transfection SystemÀº »õ·Î¿î TransIT¢ç-AAViator Transfection Reagent¿Í RevIT¢â AAV Enhancer·Î ±¸¼ºµÇ¾î ÀÖÀ¸¸ç, ÇÔ²² »ç¿ëÇÏ¿© AAV¸¦ ÃÖ´ë·Î »ý»êÇÒ ¼ö ÀÖ´Ù.
±×¸²1. Ÿ»ç Á¦Ç° ´ëºñ ³ôÀº AAV »ý»êÀ²À» º¸ÀÌ´Â TransIT¢ç-AAViator System
Viral Production 2.0 cells grown in Viral Production Medium were transfected with pAAV-WPRE-GFP, pAAV-Helper and pAAV-RepCap (AAV5, AAV8, AAV9) at a 1:1:1 plasmid concentration ratio with the following reagents: TransIT¢ç-AAViator Transfection reagent (1.25:1,vol:wt), Polymer F (1:1) , or Polymer P (1:1). RevIT¢â-AAV Enhancer (1 ¥ìl/mL) was added at the time of transfection. All AAV serotypes were harvested at 72 hours post-transfection using chemical lysis. Genome titers were determined via dPCR using primers and a probe targeting the CMV promoter region. Total assembled capsids were determined using the AAV9, AAV8 or AAV5 Titration ELISA Kits, and the Percent Full Capsids were determined by dividing the number of genome copies by total assembled capsids for each condition. The error bars represent the standard deviation of duplicate wells.)
±×¸²2. Plasmid DNA »ç¿ë·®¿¡ °ü°è¾øÀÌ ³ôÀº ¼öÀ²·Î AAV¸¦ »ý»êÇÏ´ÂTransIT-AAViator System
Viral Production 2.0 cells grown in Viral Production Medium were transfected with pAAV-WPRE-GFP, pAAV-Helper and pAAV-RepCap at a 1:1:1 plasmid concentration ratio with TransIT¢ç-AAViator Transfection reagent (1.25:1,vol:wt). RevIT¢â-AAV Enhancer (1 §¡/§¢) was added at the time of transfection. AAV was harvested at 72 hours post-transfection using chemical lysis. Genome titers were determined via dPCR using primers and a probe targeting the CMV promoter region. The error bars represent the standard deviation of duplicate wells.
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TransIT¢ç-AAViator Transfection Reagent / RevIT¢â AAV Enhancer: -10~-30¡É º¸Á¸
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