Lentivirus º¤ÅÍ ¼±Åð¡À̵å
´ç»ç´Â ´Ù¾çÇÑ application¿¡ Àû¿ë °¡´ÉÇÑ ´Ù¾çÇÑ lentivirus expression vector¸¦ Á¦°øÇÏ°í ÀÖ´Ù. ÀÌ º¤Å͵éÀº Lenti-X¢â ½Ã¸®ÁîÀÇ ½Ã½ºÅÛ¿¡¼ °¡Àå ÇÙ½ÉÀûÀÎ Á¦Ç°À¸·Î ´ëºÎºÐÀÇ mammalian cell type¿¡ Àû¿ëÀÌ °¡´ÉÇÏ´Ù (dividing ¹× non dividing cell, primary cell, stem cell, neuron Æ÷ÇÔ). ÀÌ·¯ÇÑ Lenti-X¢â ½Ã¸®Áî¿¡ lentivirus expression vectorÀÎ pLVX ½Ã¸®Áî´Â HIV-1 LTR°ú lentiviral packaging signal (¥×)À» °¡Áö°í ÀÖ°í ÀÌ¿Ü¿¡µµ transgeneÀÇ ¹ßÇö·®, ¹ÙÀÌ·¯½º ¿ª°¡, ¹ÙÀÌ·¯½ºÀÇ functionality¸¦ ³ôÀ̱â À§ÇÑ ´Ù¾çÇÑ ¼¿µéÀÌ Ãß°¡µÇ¾î ÀÖ´Ù.
- WPRE: woodchuck hepatitis virus posttranscriptional regulatory elementÀÇ ¾àÀÚ·Î poly(A) ¼¿ÀÇ readthrough¸¦ ¹æÁöÇÏ°í, RNAÀÇ processing¹× maturation °úÁ¤¿¡ ±â¿©ÇÏ¿© nuclear export¸¦ ÃËÁøÇÑ´Ù. Genomic transcript¿¡¼ ÀÌ ¼¿ÀÌ expression vectorÀÇ packagingÀ» ÃËÁøÇÏ¿© °í¿ª°¡ÀÇ lentivirus »ý»êÇϴµ¥ ±â¿©ÇÏ°í, lentivirusÀÇ transduction½Ã¿¡´Â ÀÌ ¼¿ÀÌ transgeneÀÇ ¹ßÇö¿¡ ±â¿©ÇÏ°Ô µÈ´Ù.
- cPPT/CTS: central polypurine tract/central termination sequenceÀÇ ¾àÀÚ·Î ¡°DNA flap¡±À» Çü¼ºÇÏ´Â ¼¿ÀÌ´Ù. ¡°DNA flap¡±Àº ¹ÙÀÌ·¯½ºÀÇ transduction ½Ã viral genomeÀÇ Å¸°Ù ¼¼Æ÷·ÎÀÇ nuclear import¸¦ ÃËÁøÇÏ¿© integration°ú transduction È¿À²À» ³ôÀÌ°Ô µÈ´Ù.
- RRE: Rev response elementÀÇ ¾àÀÚ·Î unspliced viral genomic RNAÀÇ nuclear export¸¦ ÃËÁøÇÏ¿© °í¿ª°¡ ¹ÙÀÌ·¯½º »ý»êÀÌ °¡´ÉÇϵµ·Ï ÇÑ´Ù.
¾Æ·¡ÀÇ lentivirus expression vector´Â ´ç»çÀÇ 4¼¼´ë Lentivirus »ý»ê ½Ã½ºÅÛÀÎ
Lenti-X¢â Packaging Single Shots (Code 631275)À» »ç¿ëÇÏ´Â °æ¿ì °£´ÜÇÑ ÇÁ·ÎÅäÄÝ·Î ¾ÈÀüÇÏ°í ½±°Ô °í¿ª°¡ÀÇ lentivirus¸¦ »ý»êÇÒ ¼ö ÀÖ´Ù.
¡á CMV ÇÁ·Î¸ðÅÍ·Î ¸ñÀû À¯ÀüÀÚ¸¦ ¹ßÇöÇÏ´Â lentivirus expression vector
ÀÌ º¤ÅÍ´Â °·ÂÇÑ constitutive promoterÀÎ CMV ÇÁ·Î¸ðÅÍ·Î lentivirus ¾È¿¡ ÀÖ´Â ¸ñÀû À¯ÀüÀÚ¸¦ ¹ßÇöÇÏ´Â º¤ÅÍÀÌ´Ù.
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Lentivirus CMV ÇÁ·Î¸ðÅÍ º¤ÅÍ ½Ã½ºÅÛ º¸·¯°¡±â
¡á EF1¥á ÇÁ·Î¸ðÅÍ·Î ¸ñÀû À¯ÀüÀÚ¸¦ ¹ßÇöÇÏ´Â lentivirus expression vector
ÀÌ º¤ÅÍ´Â CMV promoter°¡ silenceµÇ´Â ÀϺΠcell type (e.g. hematopoietic cell, stem cell µî)¿¡ Àû¿ëÇϱâ À§ÇÑ EF1¥á ÇÁ·Î¸ðÅÍÀÇ lentivirus ½Ã½ºÅÛÀÌ´Ù.
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Lentivirus EF1¥á ÇÁ·Î¸ðÅÍ º¤ÅÍ ½Ã½ºÅÛ º¸·¯°¡±â
¡á Tet-On¢ç 3G¸¦ »ç¿ëÇÑ inducible expression¿ë lentivirus expression vector
ÀÌ º¤ÅÍ´Â TRE3G promoter¸¦ »ç¿ëÇØ lentivirus ¾È¿¡ ÀÖ´Â ¸ñÀû À¯ÀüÀÚ¸¦ ¿øÇÏ´Â ½ÃÁ¡¿¡¼ ¸Å¿ì °í°¨µµ·Î inducible expressionÇϴ°ÍÀÌ °¡´ÉÇÏ´Ù. Regulator Vector¿Í Response Vector¸¦ º°µµ·Î ÁغñÇÏ´Â 2 º¤ÅÍ ½Ã½ºÅÛ°ú all-in-one º¤ÅÍ ½Ã½ºÅÛ µÎ Á¾·ù·Î ÆǸŵǰí ÀÖ´Ù.
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Lentivirus, Tet-on¢ç inducible Expression (Two vectors)
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Lentivirus, Tet One¢â Inducible Expression (One vector)
¡á ProteinÀÇ stability¸¦ Á¶ÀýÇØ ¹ßÇö·®À» Á¶ÀýÇÏ´Â ProteoTuner¸¦ »ç¿ëÇÑ lentivirus expression vector
ÀÌ º¤ÅÍÀÇ ProteoTuner destabilization domain (DD)ÀÇ downstream¿¡ ÀÖ´Â MCS¿¡ ¸ñÀû À¯ÀüÀÚ¸¦ cloning Çϸé ÇØ´ç À¯ÀüÀÚÀÇ ´Ü¹éÁúÀÇ stability¸¦
Shield 1 (Code 632189)À¸·Î Á¶ÀýÇÒ ¼ö ÀÖ´Ù. À̸¦ ÅëÇØ ¸ñÀû ´Ü¹éÁúÀÇ ¹ßÇö ¼öÁØÀ» Á¤¹ÐÇÏ°í ½Å¼ÓÇÏ°Ô Á¶ÀýÀÌ °¡´ÉÇÏ´Ù.
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Lentivirus¸¦ Æ÷ÇÔÇÑ ´Ù¾çÇÑ ¹æ¹ýÀÇ ProteoTuner System º¸·¯°¡±â
¡á Çü±¤ ´Ü¹éÁúÀ» ¹ßÇöÇÏ´Â lentivirus expression vector
ÀÌ º¤ÅÍ´Â lentivirus ¾È¿¡ Çü±¤ ´Ü¹éÁú°ú ¸ñÀûÀ¯ÀüÀÚ¸¦ °°ÀÌ ¹ßÇö (fusion protein)Çϱ⠶§¹®¿¡ ÇØ´ç À¯ÀüÀÚÀÇ ¹ßÇö°ú ÇÔ²² subcellular localizationÀ» °°ÀÌ È®ÀÎÇÒ ¼ö ÀÖ´Ù.
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´Ù¾çÇÑ Çü±¤´Ü¹éÁú ¸®Æ÷Å͸¦ Æ÷ÇÔÇÑ Lentivirus expression vectors º¸·¯°¡±â
¡á Promoterless·Î Çü±¤ ´Ü¹éÁú À¯ÀüÀÚ¸¦ °¡Áø lentivirus expression vector
ÀÌ º¤ÅÍ´Â promoter ¿¬±¸¸¦ À§ÇØ ¸ñÀûÇÏ´Â promoter¸¦ Á÷Á¢ cloningÇÏ¿© Çü±¤ ´Ü¹éÁú reporterÀÇ ¹ßÇöÇϱâ À§ÇÑ º¤ÅÍÀÌ´Ù. ÇÊ¿ä¿¡ µû¶ó ´Ù¾çÇÑ Çü±¤À» °ËÃâÇÒ ¼ö ÀÖ´Ù.
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ÇÁ·Î¸ðÅÍ ¿¬±¸¸¦ À§ÇÑ promoterless lentivirus º¸·¯°¡±â