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AAV2 Delivery Tet À¯µµ¹ßÇö ½Ã½ºÅÛ

Tet-Inducible System (AAV2)

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Clontech
634310
AAVpro¢ç Tet-One Inducible Expression System (AAV2)
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  • °­·ÂÇÏ°Ô ¹ßÇöÀ» Á¶Àý ÇÒ ¼ö ÀÖ´Â Tet-On 3G systemÀ» ±â¹ÝÀ¸·Î ÇÑ all-in-one vectorÇüÅÂ
  • ³·Àº ¹é±×¶ó¿îµå¿Í ³ôÀº À¯µµ¹ßÇö È¿À²
  • 10 ng/mlÀÇ doxycycline¿¡µµ ¹ÝÀÀÇÏ´Â ¹Î°¨µµ
  • AAVpro systemÀº helper-free packaging systemÀ» »ç¿ëÇϸç, AAV Particle ÃßÃâÀ» À§ÇÑ ÃßÃâ ½Ã¾à µîÀÌ Æ÷ÇԵǾîÀÖ´Ù.
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Tet-One SystemÀÌ °³¹ßµÇ±â Àü¿¡´Â ClontechÀÇ Tet-on, Tet-off systemÀº response vector¿Í regulatory vector(doxycycline-responsive transactivator protein)ÀÎ 2°³ÀÇ vector¸¦ À¯ÀüÀÚ ¹ßÇöÀ» Á¶ÀýÇß´Ù.

Tet-One SystemÀº ÇϳªÀÇ º¤ÅÍ¿¡ ±¸¼º¿ä¼Ò¸¦ ¸ðµÎ Æ÷ÇÔÇÏ°í ÀÖ´Ù. Tet-On 3G transactivator´Â human phosphoglycerate kinase 1 promoter¸¦ ÀÌ¿ëÇÏ¿© Á¤¹æÇâÀ¸·Î ¹ßÇöµÇ¸ç, ¹Ý´ë ¹æÇâ¿¡´Â PTRE3GS promoter¸¦ ÀÌ¿ëÇÏ¿© ¸ñÀû À¯ÀüÀÚ(Gene of interest)ÀÇ ¹ßÇöÀ» Á¶ÀýÇÑ´Ù. ±âÁ¸¿¡ ¹ßÇ¥µÈ all-in-one vector systemÀº 50~100¹è Á¤µµÀÇ À¯µµ¹ßÇö È¿À²À» º¸ÀδÙ. ÇÏÁö¸¸ ClontechÀÇ Tet-One systemÀº all-in-one vector ÇüÅ·ΠÃÖ´ë 2500¹èÀÇ À¯µµ¹ßÇö È¿À²À» º¸ÀδÙ.(Heinz et al, 2011).


±×¸². Tet-One System Mechanism - The Tet-On 3G transactivator protein is expressed constitutively from the human PGK promoter (PPGK) but is unable to bind to the TRE3G promoter (PTRE3G) in the absence of doxycycline (Dox). When bound by Dox, supplied in the culture medium, the transactivator undergoes a conformational change, binds to PTRE3G and activates transcription of a transgene cloned downstream.


±×¸² 2. Seven out of eight Tet-One clones show more than 1,000-fold induction of expression. HeLa cells were infected with LVX-TetOne-Puro-Luc lentivirus (M.O.I. = 1) and eight individual clones were selected and expanded according to the protocol. Each clone was analyzed for doxycycline-induced expression of luciferase. Seven of the eight clones demonstrated more than 1,000-fold induced expression, including two clones with 5,000-fold induction, and one clone with 10,000-fold induction.
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* AAVpro Tet-One Inducible Expression System (AAV2) (Code 634310)
AAVpro Tet-One Vector Set (Cat. No. 634311; not sold separately)
pRC2-mi342 Vector (Cat. No. 6281; not sold separately*)
pHelper Vector (Cat. No. 6282; not sold separately*)
AAVpro Extraction Solution (Cat. No. 6283; not sold separately*)
* These components are available for separate purchase in larger quantities.
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Keyword : Tet À¯µµ¹ßÇö ½Ã½ºÅÛ,Tet-inducible,Tetracycline,Å×Æ®¶ó»çÀÌŬ¸°

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