Á¦Ç°¼³¸í
º» Á¦Ç°Àº human iPS¼¼Æ÷ À¯µµ¿ë ÀçÁ¶ÇÕ retrovirus vector Á¦ÀÛÀ» À§ÇÑ plasmid DNA ÀÌ´Ù. °íÈ¿À², °í¹ßÇö À¯ÀüÀÚµµÀÔ¿ë retrovirus vectorÀÎ pDON-5 DNA (Code 3658)¿¡
iPS¼¼Æ÷ À¯µµ¿ë À¯ÀüÀÚÀÎ OCT3/4¿Í SOX2*, KLF4, LIN28°ú NANOG*ÀÌ Thosea asigna virusÀÇ2A¹è¿*1 ¸¦ »ðÀÔ ½ÃÄÑ 1°³ÀÇ º¤ÅÍ¿¡ žÀçµÇ°í ÀÖ´Ù. º» º¤ÅÍ¿Í Retrovirus
Packaging Kit Ampho (Code 6161)¿¡ Æ÷ÇԵǴ pGP Vector ¹× pE-ampho Vector¸¦ retrovirus Á¶Á¦¿ë 293 cell ¶Ç´Â 293T cell µî¿¡ co-transfection ÇÏ¿© iPS cell À¯µµ¿ë retrovirus¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù.
ÀÌ·¸°Ô Á¦ÀÛµÈ ÀçÁ¶ÇÕ retrovirus´Â RetroNectin (Recombinant Human Fibronectin Fragment) (Code T100A/B)¿¡ ´ëÇÑ Ä£È¼ºÀÌ ³ô±â ¶§¹®¿¡,
¸ñÀû¼¼Æ÷¿¡ À¯ÀüÀÚ µµÀԽà RetroNectinÀ» °íÁ¤ÈÇÑ laboratory dish¸¦ ÀÌ¿ëÇÏ´Â °ÍÀ¸·Î, ³ôÀº À¯ÀüÀÚµµÀÔ È¿À²À» ¾òÀ» ¼ö ÀÖ°í, iPS¼¼Æ÷À¯µµ È¿À²ÀÇ Çâ»óÀ» ±â´ëÇÒ ¼ö ÀÖ´Ù.
*1 5°³ÀÇ À¯ÀüÀÚ´Â ´ÜÀÏ mRNA·Î Àü»çµÇÁö¸¸, ¹ø¿ªµÉ ¶§¿¡ 2A ¹è¿ ³»ÀÇ Æ¯Á¤ÀÇ À§Ä¡¿¡¼ ºÐ¸®µÇ¾î °á°úÀûÀ¸·Î 5°³ÀÇ ´Ü¹éÁúÀÌ °ÅÀÇ ºñ½ÁÇÑ ¼öÁØÀ¸·Î ¹ßÇöÇÑ´Ù°í
»ý°¢ÇÒ ¼ö ÀÖ´Ù.
Vector´Â ´ÙÀ½°ú À¯ÀüÀÚ Á¤º¸¸¦ ¹ÙÅÁÀ¸·Î ±¸ÃàÇß´Ù.
À¯ÀüÀÚ À̸§ |
GenBank Accession No . |
OCT4 |
NM_002701.4 |
SOX2 |
NM_003106.2 |
KLF4 |
NM_004235.3*2 |
LIN28 |
NM_024674.4 |
NANOG |
NM_024865.2 |
*2 ÇöÀç´Â NM_004235.4·Î °»½ÅµÇ¾î ÀÖ¾î CDSÀÇ 5¡¯Ãø¿¡ 9 ¾Æ¹Ì³ë»ê¿¡ »ó´çÇÏ´Â ¹è¿ÀÌ Ãß°¡µÇ¾ú´Ù. º» Á¦Ç°ÀÇ KFL4 À¯ÀüÀÚ´Â N¸»´ÜÀÇ 9¾Æ¹Ì³ë»êÀ» Æ÷ÇÔÇÏÁö ¾ÊÁö¸¸,
iPS ¼¼Æ÷ À¯µµ¿¡ À¯È¿ÇÑ °ÍÀ» È®ÀÎÇÏ¿´´Ù.
Ư¡
- OCT3/4, SOX2, KLF4, LIN28, NANOG À¯ÀüÀÚ¸¦ ÇϳªÀÇ vector¿¡ žÀç
- iPS¼¼Æ÷ À¯µµ¿ë ÀçÁ¶ÇÕ retrovirus¸¦ È¿À²ÀûÀ¸·Î Á¶Á¦
- RetroNectinÀÇ º´¿ëÀ¸·ÎiPS ¼¼Æ÷ À¯µµ È¿À²µµ ¾÷
³»¿ë
pDON-5 OKSLN DNA 0.5 ¥ìg/¥ìl |
20 §¡ |
±æÀÌ 9,954 bp
º¸Á¸ -20¡É
pDON-5 OKSN DNAÀÇ ±¸Á¶