1. Choose Retroviral expression system
   : ¸ñÀû¿¡ µû¶ó ´Ù¾çÇÑ ½Ã½ºÅÛ ¼±Åà °¡´É

  2. Clone target gene
   : °£ÆíÇϰí Á¤È®ÇÑ Cloning ½Ã½ºÅÛ Ãßõ

  • Any vector, Any insert, Anyone
  • 15ºÐ ³» cloning ¹ÝÀÀ ¿Ï·á
  • 90% ÀÌ»óÀÇ ½ÇÇè Á¤È®µµ
    : ºÒÇÊ¿äÇÑ ¿°±â Ãß°¡°¡ ¾ø´Â Seamless cloning
  • Subcloning ¾øÀÌ ´Ù¾çÇÑ application Àû¿ë
  • Çâ»óµÈ colony Çü¼º ´É·ÂÀ¸·Î, ¼º°ø È¿À² Áõ°¡

  3. Package Retrovirus
   : È¿À²ÀûÀÎ Virus »ý»êÀ» À§ÇÑ packaging system Á¦°ø

Retro-X¢â Universal Packaging System (Code 631530)
  • Expression vector¿Í packaging vector¸¦ co-transfectionÇÏ¿© ¿øÇÏ´Â tropism ¼±ÅÃ
  • 4°³ÀÇ Envelope vector (pVSV-G, pEco, pAmpho, p10A1) Á¦°ø
  • HEK 293 À¯·¡ÀÇ Àü¿ë GP2-293 packaging cell Æ÷ÇÔ (º°µµ ±¸¸Å ºÒ°¡)
Pantropic Retroviral Expression System (Code 631512)
  • pVSV-G envelop vector¸¦ ÀÌ¿ëÇÏ¿©, Pantropic Retrovirus packaging °¡´É
  • HEK 293 À¯·¡ÀÇ Àü¿ë GP2-293 packaging cell Æ÷ÇÔ (º°µµ ±¸¸Å ºÒ°¡)
  • Expression vector¿Í packaging vector¸¦ co-transfection
Retro-X¢â System (Code 631508)
  • NIH/3T3 À¯·¡ÀÇ Àü¿ë RetroPack PT67 cell line (Code 631510) Æ÷ÇÔ
  • Expression vector transfection ¸¸À¸·Î 10A1 envelopÀ» °¡Áö´Â dualtropic Retrovirus packaging ¿Ï·á

[Envelope º° °¨¿° È¿À² ºñ±³]
Envelope

VSV-G

4070A (Ampho)

gap70 (Eco)

10A1 (Dual)

Tropism

Pantropic

Amphotropic

Ecotropic

Dualtropic

Receptor (target cell)

Unknown**

RAM1 (Pit2)

mCAT-1

GALV (Pit1), RAM1 (Pit2)

Possible target cell types*

Human

+

+

-

+

Mouse

+

+

+

+

Rat

+

+

+

+

Hamster

+

+/-

-

+

Cat

+

+

-

+

Dog

+

+

-

+

Monkey

+

+

-

+

Avian

+

-

-

-

Fish

+

-

-

-

Insect

+

-

-

-


* This listing of the most commonly transduced target cell types is not intended to be exclusive.
** It appears likely that a gp96 chaperone client is responsible for binding (Bloor et al. 2009)

  4. Titrate Retrovirus
   : Á¤È®ÇÑ ½ÇÇèÀ» À§ÇÑ ´Ù¾çÇÑ Titration kit º¸À¯

Retro-X¢â qPCR Titration Kit (Code 631453)
  • All-in-one kit : MMLV ±â¹ÝÀÇ RetrovirusÀÇ Á¤Á¦ºÎÅÍ qPCR±îÁö
  • qPCRÀ» ÀÌ¿ëÇÏ¿©, 4½Ã°£ ³» ½Å¼ÓÇÏ°Ô ¹ÙÀÌ·¯½º ¿ª°¡ ÃøÁ¤
  • Virus genome copy ¼ö¸¦ ÃøÁ¤ÇÏ¿© ¿ª°¡ È®ÀÎ

  5. Concentrate Retrovirus
   : °£ÆíÇÏ°Ô »ùÇà ³» ¿ª°¡ ³óÃà

Retro-X¢â Concentrator´Â retrovirusÀÇ ¿ª°¡¸¦ ÃÖ´ë 100 ¹è±îÁö ³óÃàÇÒ ¼ö ÀÖ¾î, ´õ ¸¹Àº ¹ÙÀÌ·¯½º¸¦ ¸¸µé°Å³ª, Ãß°¡·Î packaging ¼¼Æ÷¿¡ transfectionÇÒ ÇÊ¿ä ¾øÀÌ ÃæºÐÈ÷ ³ôÀº MOI·Î ¸ñÀû¼¼Æ÷¸¦ °¨¿°½Ãų ¼ö ÀÖ´Ù.

Retro-X¢â Concentrator (Code 631455)
  • Ãʰí¼Ó¿ø½ÉºÐ¸®±â ¾øÀÌ ¹ÙÀÌ·¯½º ¿ª°¡¸¦ ÃÖ´ë 100 ¹è±îÁö Áõ°¡
  • °£´ÜÇÑ Á¶ÀÛ °úÁ¤ : Mix, wait and spin
  • ¹ø°Å·Î¿ò ¾øÀÌ ºÎÇǸ¦ ½±°Ô scale up °¡´É

  6. Transduce target cells with enhancer
   : ÃÖ°íÀÇ È¿À²À» À§ÇÑ support tool Á¦°ø

RetroNectin¢ç (Recombinant Human Fibronectin Fragment) (Code T100A)
  • ÃÖ°íÀÇ Transduction enhancer : Lentivirus, RetrovirusÀÇ À¯ÀüÀÚ µµÀÔ È¿À² ±Ø´ëÈ­
  • Low toxicity for cells : Polybrene, Protamine ´ëºñ ³·Àº µ¶¼º
  • Enhanced T cell expansion : ±âÁ¸ T cell È®´ë¹è¾ç¹ý ´ëºñ ¾ÐµµÀûÀÎ T cell È®´ë¹è¾ç °¡´É (CAR-T cell ¿¬±¸¿¡ ÃÖÀû)
  • Research grade, GMP grade ¼±Åà °¡´É

   7. Analyze integration
   : ÃÖ°íÀÇ È¿À²À» À§ÇÑ support tool Á¦°ø

Retrovirus´Â RNA genome virus·Î, DNA·Î ¿ªÀü»çµÇ¾î host genome¿¡ ¹«ÀÛÀ§·Î »ðÀԵDZ⠶§¹®¿¡ ¸ñÀûÀ¯ÀüÀÚ ¹× endogenous À¯ÀüÀÚ ¹ßÇö¿¡ ¿µÇâÀ» ÁÙ ¼ö ÀÖ´Ù. µû¶ó¼­, provirusÀÇ »ðÀÔºÎÀ§ÀÇ ½Äº°Àº host cellÀÇ Ç¥ÇöÇü ¿¬±¸¿¡ Áß¿äÇÏ´Ù.
Retro-X¢â Integration Site Analysis Kit (Code 631467)
  • GenomeWalker¸¦ ±â¹ÝÀ¸·Î provirusÀÇ °Ô³ð »ðÀÔ ºÎÀ§ È®ÀÎ
  • MMLV ¹× MSCV ŸÀÔÀÇ retroviral vector¿¡ Àû¿ë °¡´É