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Home > ÀüÁ¦Ç°º¸±â > Gene transfer (virus) > Lentivirus > 3¼¼´ë Lentivirus packaging system
ÀÓ»ó Ä¡·áÁ¦ °³¹ßÀ» À§ÇÑ

3¼¼´ë Lentivirus packaging system

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
6195
LVpro¢â Packaging Mix
°ü·ÃÇмú ±¸¸ÅÇϱâ
60 Rxns
1,163,200¿ø 
1,454,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
6195_6962-6967_e.v2202Da.pdf
Takara
6962
LVpro¢â Packaging Mix (pLVpro-MSCV Vector)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 Set
(Package)
1,387,200¿ø 
1,734,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
6195_6962-6967_e.v2202Da.pdf
Takara
6963
LVpro¢â Packaging Mix (pLVpro-MSCV-EI Vector)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 Set
1,387,200¿ø 
1,734,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
6195_6962-6967_e.v2202Da.pdf
Takara
6964
LVpro¢â Packaging Mix (pLVpro-EF1¥á Vector)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 Set
(Package)
1,387,200¿ø 
1,734,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
6195_6962-6967_e.v2202Da.pdf
Takara
6965
LVpro¢â Packaging Mix (pLVpro-MSCV-ZsGreen1 Vector)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 Set
1,387,200¿ø 
1,734,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
6195_6962-6967_e.v2202Da.pdf
Takara
6966
LVpro¢â Packaging Mix (pLVpro-MSCV-EI-ZsGreen1 Vector)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 Set
1,387,200¿ø 
1,734,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
6195_6962-6967_e.v2202Da.pdf
Takara
6967
LVpro¢â Packaging Mix (pLVpro-EF1¥á-ZsGreen1 Vector)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 Set
1,387,200¿ø 
1,734,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
6195_6962-6967_e.v2202Da.pdf

  • Tat-independentÇÑ 3¼¼´ë Lentivirus packaging system
    - HIV-1 À¯·¡ÀÇ ¼­¿­À» ÃÖ¼ÒÈ­ÇÏ¿© »ç¿ë ¾ÈÀü¼º Çâ»ó, ÀÓ»ó ¿¬±¸¿¡ ÃÖÀû
    - 5¡¯ LTR U3 ºÎÀ§°¡ CMV promoter·Î ´ëüµÇ¾î, tat trans-activator ¾øÀ̵µ virus »ý»ê °¡´É
    - 3¡¯ LTR ÀϺΠ°á¼ÕÀ¸·Î self-inactivation (SIN) µÇ¾î, ¹ÙÀÌ·¯½º ÀÚ°¡ º¹Á¦ ¹æÁö
  • ³ôÀº ¿ª°¡ (titer)ÀÇ virus »ý»êÀ» À§ÇØ ÃÖÀûÈ­µÈ ÆÐŰ¡ ÇÁ·ÎÅäÄÝ
  • ½ÇÇè¿¡ µû¸¥ ´Ù¾çÇÑ expression vector ¼±ÅÃ
    - ¸ñÀû À¯ÀüÀÚ¿¡ µû¸¥ ÃÖÀûÀÇ promoter ¼±Åà °¡´É
    (1) Promoter ¼±Åà (MSCV, MSCV-EI, EF1¥á)
    (2) Çü±¤ ´Ü¹éÁú À¯¹« ¼±Åà (ZsGreen1)
Á¦Ç° ¼³¸í
LVpro Packaging Mix´Â ¿©·¯ °³ÀÇ packaging plasmid (env, gag, pol, rev ¹ßÇö)°¡ premix ÇüÅ·ΠÁ¦°øÇϸç, ¿¬±¸ÀÚ°¡ ½ÇÇè Á¶°Ç ÃÖÀûÈ­ ¹× ¼ÂÆà °úÁ¤À» °ÅÄ¡Áö ¾Ê°íµµ ³ôÀº ¿ª°¡ÀÇ Lentivirus¸¦ »ý»êÇÒ ¼ö ÀÖ´Ù. º» Á¦Ç°Àº HIV-1 À¯·¡ÀÇ tat ¼­¿­À» Á¦°ÅÇÏ°í 5¡¯ LTRÀÇ U3 ºÎÀ§´Â CMV promoter·Î ´ëüÇÏ¿© tat-independentÇÑ lentivirus¸¦ »ý»êÇÒ ¼ö ÀÖ´Â pLVpro lentiviral vector plasmidÀ» packaging cell¿¡ co-transfectionÇÏ´Â °ÍÀ¸·Î ½ÇÇèÀ» ÁøÇàÇÑ´Ù. pLVpro lentiviral vector plasmid¿Í LVpro Packaging Mix¸¦ »ç¿ëÇØ »ý»êµÈ ¹ÙÀÌ·¯½º´Â HIV-1 À¯·¡ÀÇ Gag, Pol, Rev ´Ü¹éÁú°ú VSV-G envelopeÀ» ¹ßÇöÇÑ´Ù.

±×¸² 1. Lentivirus »ý»ê workflow



¹ÙÀÌ·¯½º Á¦ÀÛÀ» À§ÇÑ transfection ½Ã¿¡´Â º¸´Ù ³ôÀº ¿ª°¡ÀÇ ¹ÙÀÌ·¯½º¸¦ »ý»êÇϵµ·Ï ±¸ÃàµÈ Lenti-X¢â 293T Cell Line (Code 632180)°ú functionalÇÑ ¹ÙÀÌ·¯½º »ý»ê È¿À²À» ³ô¿©ÁÖ´Â TransIT¢ç-VirusGEN¢â Transfection Reagent (Code MIR 6700)À» ÃßõÇÑ´Ù.

¹ÙÀÌ·¯½º »ý»ê È¿À²À» ºñ±³Çϱâ À§ÇØ, A»çÀÇ packaging plasmid¿Í transfection ½Ã¾à, ´ç»çÀÇ LVpro Packaging Mix (Code 6195)¿Í Ãßõ ½Ã¾à TransIT¢ç-VirusGEN¢ç Transfection Reagent (Code MIR 6700)À» »ç¿ëÇÏ¿´´Ù. ±× °á°ú, ´ç»ç ½Ã½ºÅÛÀ» ÀÌ¿ëÇßÀ» ¶§ °¡Àå ³ôÀº »ý»ê ¿ª°¡¸¦ º¸¿´´Ù.

Ç¥1. Ÿ»ç packaging system°úÀÇ »ý»ê È¿À² ºñ±³¸¦ À§ÇÑ ½ÇÇè µðÀÚÀÎ
* Company A¡¯s vector´Â CMV promoter ÇÏ·ù¿¡¼­ ZsGreen1 ¹ßÇö



±×¸²1. LVpro Packaging Mix (Code 6195)¿Í Ÿ»ç Á¦Ç° °£ÀÇ ¹ÙÀÌ·¯½º »ý»ê È¿À² ºñ±³
HT-1080 ¼¼Æ÷¿¡¼­ Å×½ºÆ®ÇÑ °á°ú, Ÿ»çÀÇ packaging ½Ã½ºÅÛÀ» ÀÌ¿ëÇÑ 1¹ø ½ÇÇè ±º ´ëºñ ´ç»ç packaging ½Ã½ºÅÛÀ» »ç¿ëÇÑ 4¹ø ½ÇÇè ±ºÀÌ 10¹è ÀÌ»óÀÇ ³ôÀº »ý»ê È¿À²À» º¸¿´´Ù.



Expression vector ±¸Á¶
- SIN 5¡®LTR: CMV promoter hybrid 5¡¯ LTR (U3 free, tat independent)
- Lentivirus packaging signal (¥×)
- Minimal gag sequence
- RRE (Rev response element)
- cPPT/CTS (central polypurine tract/central termination sequence)
- GOI ¹ßÇö promoter
   Option 1) MSCV promoter: ¸ñÀû À¯ÀüÀÚ°¡ Å©°Å³ª, ¾ÈÁ¤ÀûÀÎ ¹ÙÀÌ·¯½º »ý»ê ¿ª°¡°¡ ¿ä±¸µÉ ¶§
   Option 2) MSCV-EI promoter: Å©±â°¡ Å« ¸ñÀû À¯ÀüÀÚÀÇ °ú¹ßÇöÀ» À¯µµÇÏ°íÀÚ ÇÒ ¶§
   (Human EF1¥á À¯ÀüÀÚ À¯·¡ÀÇ exon, intron ¼­¿­À» MSCV-U3 promoter ÇÏ·ù¿¡¼­ ¹ßÇö)
   (´Ü, ¸ñÀû À¯ÀüÀÚÀÇ Å©±â¿Í ¼­¿­¿¡ µû¶ó, MSCV promoter¿¡ ºñÇØ »ý»ê ¿ª°¡°¡ ³·À» ¼ö ÀÖÀ½)
   Option 3) EF1¥á promoter: ¸ñÀû À¯ÀüÀÚÀÇ °ú¹ßÇöÀ» À¯µµÇÏ°íÀÚ ÇÒ ¶§
- WPRE (woodchuck hepatitis virus post-transcriptional regulatory element): X proteinÀÌ ³·°Ô ¹ßÇöÇϵµ·Ï °³·®µÇ¾î, transductionµÈ ¸ñÀû ¼¼Æ÷¿¡¼­ÀÇ tumorigenesis À¯¹ß È®·ü °¨¼Ò
- SIN 3¡®LTR (U3 free)

* Vector mapÀ» Ŭ¸¯Çϸé, vector sequence fasta ÆÄÀÏ (txt. È®ÀåÀÚ)¸¦ È®ÀÎÇÒ ¼ö ÀÖ½À´Ï´Ù.

MSCV promoter

pLVpro-MSCV Vector
(#6962 ±¸¼ºÇ°)

pLVpro-MSCV-ZsGreen1 Vector
(#6965 ±¸¼ºÇ°)

MSCV-EI promoter

pLVpro-MSCV-EI Vector
(#6963 ±¸¼ºÇ°)
pLVpro-MSCV-EF1-ZsGreen1 Vector
(#6966 ±¸¼ºÇ°)

EF1¥á promoter

pLVpro-EF1 ¥á Vector
(#6964 ±¸¼ºÇ°)
pLVpro-EF1 ¥á -ZsGreen1 Vector
(#6967 ±¸¼ºÇ°)






Keyword : Lentivirus,stable cell ±¸Ãà,stable cell line,CAR-T,Ä¡·áÁ¦,therapeutics,CAR,¼¼Æ÷Ä¡·áÁ¦,ÀÓ»ó,3¼¼´ë Lentivirus,3rd generation,tat-independent,tat free,U3 free,SIN,Self inactivation,self inactivated

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Tet-on inducible Expression (Two vectors)
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Lenti-X¢â p24 Rapid Titer Kit (Single Wash)
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Ecotropic Receptor Booster
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