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Regulated Transcription System

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
635081
iDimerize Regulated Transcription System
°ü·ÃÇмú ±¸¸ÅÇϱâ
Each
2,384,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32733, x33388, x33618, x33619

  • iDimerize ±â¼úÀ» ÀÌ¿ëÇÏ¿© Àü»ç¸¦ °­·ÂÇÏ°Ô Á¶Àý
  • Àü»ç¸¦ Á¶ÀýÇÏ´Â ÀÛÀº ºÐÀÚ ligandÀÇ ÅõÀÔ¿¡ µû¶ó Á¶Àý °¡´É
  • Ligand¸¦ ÅõÀÔÇϱâ Àü¿¡´Â È°¼ºµµ¸ÞÀÎÀÌ DNA·ÎºÎÅÍ ºÐ¸®µÇ¾î Àֱ⠶§¹®¿¡ ¹é±×¶ó¿îµå°¡ ¸Å¿ì ³·À½
  • A/C Heterodimerizer´Â AP21967 ligand¿Í µ¿ÀÏÇÔ
iDimerize ±â¼úÀº ¸ñÀû À¯ÀüÀÚÀÇ Àü»ç È°¼ºÀ» ¸Å¿ì °­·ÂÇÏ°Ô Á¶ÀýÇÒ ¼ö ÀÖ´Ù. ƯÈ÷ Àü»ç Á¶Àý¿¡ À־ iDimerize ±â¼úÀÇ °¡Àå Áß¿äÇÑ ÀåÁ¡ Áß Çϳª´Â dimerizer ligand°¡ ÷°¡µÇ±â Àü¿¡ Àü»çÈ°¼º µµ¸ÞÀÎ(AD)°¡ DNA¿Í ºÐ¸®µÇ¾î Àֱ⠶§¹®¿¡ ¹é±×¶ó¿îµå°¡ ³·´Ù´Â Á¡ÀÌ´Ù. Àü»ç ¿ä¼ÒµéÀº µÎ °¡Áö ±â´ÉÀÇ ´Ü¹éÁúµé·Î, DNA binding domainÀ» ÅëÇØ ¸ñÀûÀ¯ÀüÀÚ ±ÙóÀÇ Æ¯ÀÌÀû DNA¼­¿­À» ÀÎÁöÇÑ ÈÄ transcription activation domainÀ» ÅëÇØ Àü»ç¸¦ È°¼ºÈ­ ½ÃÅ°µµ·Ï ¼¼Æ÷ÀÇ Àü»ç ±âÀÛÀ» ¸ðÀ¸´Â ¿ªÇÒÀ» ÇÑ´Ù. ÀÌµé µÎ µµ¸ÞÀÎÀº °¢ÀÚ ´Ü¹éÁúÀÌ ¹ßÇöµÇ¸é A/C Heterodimerizer ligand¿¡ ÀÇÇØ ¼­·Î dimerizationÀ» ¸¸µé¾î Àü»ç¸¦ È°¼ºÈ­½Ãų ¼ö ÀÖ´Ù.
A/C Heterodimerizer Ligand (AP21967)
A/C Heterodimerizer´Â ÇÕ¼ºµÈ ¼¼Æ÷Åõ°ú¼º ligand·Î µÎ °³ÀÇ À¶ÇմܹéÁúÀÇ heterodimerizationÀ» À¯µµÇϴµ¥ »ç¿ëµÈ´Ù. ¿¹¸¦ µé¸é, ÀÌ Å°Æ®¸¦ »ç¿ëÇϸé DmrC-tagged transcription activation domainÀº DmrA-tagged DNA binding domain°úÀÇ dimerize¸¦ À¯µµÇÒ ¼ö ÀÖ´Ù.
Regulating Transcription
¼¼Æ÷·Î ¾Æ·¡ Çö󽺹̵带 ÇüÁúÀüȯ ½ÃŲ´Ù
1. transcription activation°ú DNA binding components¸¦ ¹ßÇöÇÏ´Â Çö󽺹̵å(pHet-Act1-2)
2. PZI-1 inducible promoter(in pZFHD1-2)ÀÇ ÇÏ·ù¿¡ Ŭ·Î´×µÈ ¸ñÀûÀ¯ÀüÀÚ

DNA binding component´Â promoter¿¡ °áÇյǾî ÀÖ¾î DmrA¿Í DmrC¸¦ ÅëÇØ Àü»çÈ°¼º¿ä¼ÒµéÀÌ °áÇÕÇÏ¿© Àü»ç°¡ È°¼ºÈ­ µÉ ¶§±îÁö Àü»ç¸¦ ¾ïÁ¦ÇÑ´Ù. A/C Heterodimerizer°¡ ÷°¡µÇ¸é µÎ components°¡ °áÇÕÇÏ°í PZI-1 promoter·ÎºÎÅÍ ¸ñÀû À¯ÀüÀÚ°¡ Àü»çµÈ´Ù.



The iDimerize Regulated Transcription System uses the tight PZI-1 promoter, which exhibits very low background in the absence of induction because it is not recognized by host transcription factors. A further level of control is provided separating the DNA binding component (DmrA/DB; green) and transcription activation component (DmrC/TA; yellow) into separate compartments in the cell. DmrC/TA does not possess a nuclear localization signal (NLS), so it resides predominantly in the cytoplasm; however, it is brought to the nucleus by the addition of A/C heterodimerizer which causes its interaction with DmrA/DB. DmrA/DB does possess an NLS, so when A/C heterodimerizer is added, the transcription factor is completed through dimerization. The complete transcription factor is translocated into the nucleus, where it binds the PZI-1 promoter and induces expression of the gene of interest.



Tightly regulated transcription using iDimerize technology. Clone your gene of interest downstream of the ZHFD1 inducible promoter (PZI-1). The DNA binding component (DmrC/DNA-BD fusion; red) recognizes and binds sequences within the promoter. However, activation of transcription only occurs when the DmrC/DNA-BD dimerizes with the transcription activation component (DmrA-AD fusion; green) at the promoter, when the DmrA and DmrC domains both bind to the A/C Heterodimerizer (AP21967).



Regulated transcription of luciferase using the iDimerize Regulated Transcription System. Firefly luciferase was cloned into the multiple cloning site of the pZFHD1-2 vector. The plasmid was then cotransfected into HEK 293 cells with pHet-Act1-2 (which expresses the transcription activation components). Cells were treated with the indicated doses of A/C Heterodimerizer, and induced gene expression was measured 24 hr later. Induced expression of luciferase was measured using a luminometer. RLU = Relative Light Units.



Dose-dependent control of gene expression with the iDimerize Inducible Expression System. HT1080 cells were stably transfected with the secreted alkaline phosphatase (SEAP) reporter gene and the DmrC-AD/DmrA-DBD constructs, and treated with or without A/C Heterodimerizer. In the absence of A/C Heterodimerizer, target gene expression was undetectable. Half-maximal induction occurred at 2 nM A/C Heterodimerizer.

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