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Gene transfer (virus)
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Adenovirus
> Adeno-X¢â workflow
Adeno-X¢â workflow
1. Choose Adenoviral expression system
: ¸ñÀû¿¡ µû¶ó ´Ù¾çÇÑ ½Ã½ºÅÛ ¼±Åà °¡´É
2. Clone target gene
: °£ÆíÇÏ°í 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 Adenovirus
: È¿À²ÀûÀÎ Virus »ý»êÀ» À§ÇÑ packaging cell Á¦°ø
Adeno-X¢â 293 Cell Line (Code 632271)
±âÁ¸ÀÇ strainº¸´Ù °ÇÑ ºÎÂø·Â(adherent)
¼ºÀå ¼Óµµ°¡ ´À¸®±â ¶§¹®¿¡ º¸´Ù È¿À²ÀûÀ¸·Î adenovirus Á¦ÀÛ ¹× ÁõÆø °¡´É
4. Titrate Adenovirus
: Á¤È®ÇÑ ½ÇÇèÀ» À§ÇÑ ´Ù¾çÇÑ titration kit º¸À¯
ÀçÇö¼º ÀÖ´Â ½ÇÇè °á°ú¸¦ ¾ò±â À§Çؼ´Â Á¤È®ÇÑ ¹ÙÀÌ·¯½º ¿ª°¡¸¦ ÃøÁ¤ÇÏ´Â °ÍÀÌ ÇʼöÀûÀÌ´Ù. Adeno-X¢â½Ã¸®Áî´Â ±âÁ¸ ¹æ¹ý (Plaque assay, End-point dilution assay)º¸´Ù Á¤È®ÇÏ°í ºü¸£°Ô Adenovirus ¿ª°¡¸¦ ÃøÁ¤ÇÒ ¼ö ÀÖ´Ù.
5. Purify Adenovirus
: °£ÆíÇÏ°í È¿À¯ÀûÀÎ Adenovirus ÃßÃâ kit Á¦°ø
°£´ÜÇÑ Á¶ÀÛÀ¸·Î 90ºÐ ³», °í¿ª°¡, °í¼øµµÀÇ Adenovirus Á¤Á¦
CsClÀ̳ª ÃÊ°í¼Ó¿ø½ÉºÐ¸®±â°¡ ºÒÇÊ¿äÇÑ chromatograpy Á¤Á¦
Overview of the Adeno-X¢â Maxi Purification protocol
6. Transduce target cells with enhancer
: ÃÖ°íÀÇ È¿À²À» À§ÇÑ support tool Á¦°ø
CAR Receptor Booster (Code 631470)
ÀϽÃÀûÀ¸·Î ¸ñÀû ¼¼Æ÷ÀÇ Adenoviral receptorÀÇ ³óµµ¸¦ Áõ°¡½ÃÄÑ, transduction È¿À² Áõ°¡
CAR receptor°¡ Àû´Ù°í ¾Ë·ÁÁø hematopoietic cell, CHO cellÀ» Æ÷ÇÔÇÑ ´ëºÎºÐÀÇ cell¿¡ ¹ü¿ëÀûÀ¸·Î »ç¿ë °¡´É