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Gene transfer (virus)
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Lentivirus
> Lenti-X¢â Transduction Sponge
½Å°³³ä lentivirus transduction
Lenti-X¢â Transduction Sponge
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Clontech
631478
Lenti-X¢â Transduction Sponge
24 ȸ
995,000¿ø
Product index
Adeno-Associated Virus (AA..
Overview - All about A..
Packaging (293T Cell L..
AAV1, 2, 5, 6 Vector S..
CRE Recombinase System
LacZ System
pAAV-ZsGreen1 Vector
CRISPR/Cas9 Vector Sys..
CRISPR/SaCas9 Vector S..
Tet-Inducible System (..
Cell & Sup. Purificati..
Purification Kit Maxi/..
Purification Kit (AAV2..
AAVpro¢ç Concentrator
AAVpro¢ç Titration Kit..
Extraction Solution
AAV Freeze-thaw extrac..
Lentivirus
[°³¿ä] Packaging Singl..
[FAQ] Lentiviral Gene ..
CMV promoter º¤ÅÍ ½Ã½º..
EF1 ¥á promoter º¤ÅÍ ..
Lenti-X¢â SARS-CoV-2 P..
3¼¼´ë Lentivirus packa..
Çü±¤´Ü¹éÁú ¸®Æ÷ÅÍ º¤ÅÍ
Tet-on inducible Expre..
Tet One Inducible Expr..
Lenti-X¢â CRISPR/Cas9 ..
(Legacy) Tet-Inducible..
Reporter assay ½Ã½ºÅÛ ..
Packaging single shots
Packaging Cell Lines
Lenti-X¢â qRT-PCR Titr..
Lenti-X¢â p24 Rapid Ti..
GoStix¢â Plus
Lentivirus ³óÃà ½Ã¾à
Lentivirus Á¤Á¦
Ecotropic Receptor Boo..
RetroNectin¢ç (Recombi..
Transduction ½Ã¾à
Provirus Quantitation ..
Integration Site Analy..
Lenti-X¢â Transduction..
TransIT¢ç Lentivirus S..
Retrovirus
Retro-X ¼±Åà °¡À̵å
RetroNectin¢çÀÇ ´Ù¾çÇÑ..
Packaging Cell £¦ ½Ã½º..
Retrovirus vector £¦ ..
Çü±¤´Ü¹éÁú Retrovirus ..
MSCV Retrovirus ½Ã½ºÅÛ
Retro-X Q vector
pDON-5 DNA
pDON-AI-2 DNA
pMEI-5 DNA
Retrovirus Packaging K..
Tet À¯µµ¹ßÇö ½Ã½ºÅÛ (T..
Tet One Inducible Syst..
qRT-PCR titer ÃøÁ¤
Titer Set (for Real Ti..
Retrovirus ³óÃà
Receptor Boosters (Eco..
Provirus Copy ¼ö ÃøÁ¤
Integration Site Analy..
Adenovirus
Adeno-X¢â workflow
Adenoviral Expression ..
Packaging Cell Lines
Adenovirus Á¤Á¦ (Maxi ..
Adenovirus Á¤Á¦ (Mega ..
qPCR titer ÃøÁ¤ Kit
Hexon ±â¹Ý titer ÃøÁ¤
Adeno-X GoStix
CAR Receptor Booster
Mammalian Cell ¿ë Vector
[¼±Åð¡À̵å] ´Ù¾çÇÑ Á¾..
¾ç¹æÇ⼺ ¹ßÇö º¤ÅÍ (Bi..
EF1¥á Promotor Vector ..
epitope tag (HA or c-M..
in vivo Vector, pLIVE ..
Magnetic Myc IP Kit
pBApo-CMVvector ½Ã¸®Áî
pBApo-EF1¥á Vector ½Ã..
pIRES Bicistronic Expr..
´Ù°ø¼º ½ºÆÝÁö·Î ºü¸£°í È¿°úÀûÀ¸·Î lentivirus transduction È¿À² Áõ´ë
½ºÆÝÁö Çϳª·Î 24 well plate Å©±âÀÇ 1 well¿¡¼ 10
5
~10
7
cell¿¡ transduction °¡´É
Spinoculation°ú polybreneÀ» »ç¿ëÇÏÁö ¾Ê¾Æ ¼¼Æ÷ ¼Õ»óÀÌ ÀûÀ½
TransductionÀÌ ¾î·Á¿î primary cell ¹× ´Ù¾çÇÑ cell-line¿¡ Àû¿ë °¡´É
ÀÛÀº reaction volumeÀ¸·Î ¼Ò·®ÀÇ ¹ÙÀÌ·¯½º·Îµµ ½ÇÇè °¡´É
Á¦Ç°¼³¸í
Lenti-X¢â Transduction Sponge´Â ´Ù°ø¼º alginate (¾Ë±ä»ê)·Î ±¸¼ºµÈ ½ºÆÝÁö·Î °£ÆíÇÏ°Ô lentivirus transductionÀ» ÃËÁøÇÑ´Ù. ½ºÆÝÁöÀÇ ´Ù°ø¼º microfluidics ±¸Á¶´Â ¼¼Æ÷¿Í lentivirus°£ ¹°¸®Àû °Å¸®¸¦ Á¼Çô spinoculationÀ̳ª chemical enhancer (e.g. Polybrene) ¾øÀ̵µ ¼¼Æ÷ µ¶¼ºÀ» ÃÖ¼ÒÈÇÏ¸é¼ ºü¸¥ ½Ã°£ ¾È¿¡ È¿°úÀûÀ¸·Î transductionÀ» ÁøÇàÇÑ´Ù. 24 well plate Å©±âÀÇ ½ºÆÝÁö´ç Çѹø¿¡ 10
5
~10
7
cellÀ» transductionÇÒ ¼ö ÀÖÀ¸¸ç º°µµÀÇ microfluidics ±â±â ¾øÀÌ ÇüÁúµµÀÔÀÌ ¾î·Á¿î cell-line (ºÎÂøÇü, ºÎÀ¯Çü ¼¼Æ÷ ¸ðµÎ °¡´É) ¹× primary cell µî ´Ù¾çÇÑ ¼¼Æ÷¿¡ transductionÀ» ÁøÇàÇÒ ¼ö ÀÖ´Ù.
cell handlingÀÌ ÂªÀº °£ÆíÇÑ workflow, Mix-Drop-Spin
±×¸² 1. Lenti-X¢â Transduction Sponge workflow
1. Ÿ°Ù ¼¼Æ÷¿Í lentivirus¸¦ È¥ÇÕ (ÃÑ ºÎÇÇ ÃÖ´ë 150 §¡).
2. ½ºÆÝÁö¸¦ 24well plate·Î well´ç 1°³¾¿ ¿Å±â°í 1¹øÀÇ mixture¸¦ ½ºÆÝÁö À§·Î dropwise ¹æ½ÄÀ¸·Î ÷°¡ ¹× Èí¼ö.
3. 37 ¡ÉÀÇ CO
2
ÀÎÅ¥º£ÀÌÅÍ¿¡¼ 1½Ã°£µ¿¾È incubation ÈÄ well¿¡ ¹èÁö¸¦ Ãß°¡·Î 1 §¢ ÷°¡, ÀÌÈÄ ÀÎÅ¥º£ÀÌÅÍ¿¡¼ 16~24½Ã°£ ¹è¾ç.
4. ´ÙÀ½³¯ ¹èÁö¿Í ½ºÆÝÁö¸¦ ¸ðµÎ 15 §¢ Æ©ºê·Î ¿Å±â°í Release Buffer 1 §¢¸¦ ÷°¡ ÈÄ ¿ø½ÉºÐ¸®ÇÏ¿© ½ºÆÝÁö¸¦ ¿ëÇØ, ¿ëÇØ ÈÄ ¹æÃâµÈ ¼¼Æ÷¸¦ PBS·Î 2ȸ wash.
5. TransductionµÈ ¼¼Æ÷¸¦ »õ culture vessel·Î À̵¿ ÈÄ ¹è¾ç.
±×¸² 2. Lenti-X¢â Transduction Sponge¿Ü°ü ¹× »ç¿ë¹ý
A: Á¦Ç° »çÁø (°³º° Æ÷ÀåµÈ ½ºÆÝÁö 24°³)
B: ´Ù°ø¼º ½ºÆÝÁöÀÇ È®´ë»çÁø (150¹è), 20~300 §ÀÇ microfluidics ±¸Á¶
C: ÇɼÂÀ¸·Î ½ºÆÝÁö¸¦ 24 well plate·Î ¿Å±â´Â °úÁ¤
D: ½ºÆÝÁö¿¡ ¼¼Æ÷¿Í ¹ÙÀÌ·¯½º mixture¸¦ ÷°¡ÇÑ µÚ 1½Ã°£ µ¿¾È incubationÇÏ´Â °úÁ¤
ÆгΠE: Transduction (16~24½Ã°£ incubation)ÈÄ ½ºÆÝÁö¸¦ 15 §¢ Æ©ºê·Î ¿Å±â´Â °úÁ¤
½ÇÇè µ¥ÀÌÅÍ
±×¸² 3. Lenti-X¢â Transduction SpongeÀº ´Ù¾çÇÑ cell¿¡¼ spinoculation (with polybrene)°ú µ¿µîÇϰųª ´õ È¿°úÀûÀÎ transductionÀÌ °¡´É.
A: 1 X 10
6
Jurkat ¼¼Æ÷¿¡ spinoculation°ú Lenti-X¢â Transduction Sponge ¹æ¹ýÀ¸·Î ZsGreen1À» ¹ßÇöÇÏ´Â lentivirus¸¦ MOI 0, 1, 5, 10·Î transductionÇÑ °á°úÀÌ´Ù. SpinoculationÀº cell plate¸¦ Polybrene 8 §¶/§¢°ú ÇÔ²² 32¡É¿¡¼ 1,400 x g·Î 90ºÐ°£ ¿ø½ÉºÐ¸®ÇÏ¿´°í, Lenti-X¢â Transduction Sponge´Â Á¦Ç°ÀÇ »ç¿ëÀÚ¸Þ´º¾ó¿¡ µû¶ó ÁøÇàÇÏ¿´´Ù. µÎ ¹æ¹ý ¸ðµÎ transduction 48½Ã°£ µÚ ZsGreen1ÀÇ ¹ßÇöÀ²À» FCM (Flow Cytometry)À¸·Î ºÐ¼®ÇÏ¿© Á¤¸®ÇÏ¿´´Ù. ±× °á°ú Lenti-X¢â Transduction Sponge°¡ spinoculation ¹æ¹ýº¸´Ù °£ÆíÇÏ°í ´õ¿í È¿°úÀûÀ̾ú´Ù.
B: ZsGreen1 ¹ßÇö lentivirus¸¦ ÆгΠA¿Í µ¿ÀÏÇÑ ¹æ¹ýÀ¸·Î ´Ù¾çÇÑ cell line¿¡¼ transductionÇÑ °á°úÀÌ´Ù. ´Ù¾çÇÑ ¼¼Æ÷¿¡¼ Lenti-X¢â Transduction Sponge´Â spinoculation ¹æ¹ý°ú µ¿µîÇϰųª ´õ È¿°úÀûÀ̾ú´Ù.
±×¸² 4. Lenti-X¢â Transduction Sponge´Â primary T cellÀÇ viability¸¦ À¯ÁöÇÏ¸é¼ transduction È¿À²À» ³ôÀÏ ¼ö ÀÖ´Ù.
A. Human primary T cellÀ» MHC tetramer, antiCD3/CD28-coated bead, RetroNectin+CD3, CD3¸¦ °¢°¢ »ç¿ëÇÏ¿© È°¼ºÈ½ÃÄ×°í, 48½Ã°£ µÚ CD69+ÀÇ ¹ßÇöÀ» ÃøÁ¤ÇÏ¿© È°¼ºÈ Á¤µµ¸¦ ÃøÁ¤ÇÏ¿´´Ù. ÀÌÈÄ 1 x 10
6
°³ÀÇ È°¼ºÈµÈ ¼¼Æ÷¿¡ Lenti-X¢â Transduction Sponge¸¦ »ç¿ëÇØ 24½Ã°£ µ¿¾È ZsGreen1À» ¹ßÇöÇÏ´Â lentivirus·Î transductionÇÏ°í, transduction 48½Ã°£ ÈÄ FACS·Î ZsGreen1ÀÇ ¹ßÇöÀ²À» ÃøÁ¤ÇÏ¿© %·Î Á¤¸®ÇÑ °á°úÀÌ´Ù.
B. Transduction 48½Ã°£ ÈÄ 7-AAD ¿°»ö°ú FACS¸¦ »ç¿ëÇØ viability¸¦ ÃøÁ¤ÇÑ °á°ú, ³ôÀº viability¸¦ È®ÀÎÇÒ ¼ö ÀÖ¾ú´Ù.
º¸Á¸: 4¡É (Lenti-X¢â Transduction Sponges´Â °ÇÁ¶Á¦¿Í ÇÔ²² ÆÄ¿ìÄ¡ º¸°ü)
³»¿ë
Lenti-X¢â Transduction Sponges
24
Release Buffer
30 §¢
Forceps
1
°ü·ÃÁ¦Ç°
- CGT ¿¬±¸¿¡ ÁÖ·Î »ç¿ëµÇ´Â cellµé¿¡ Àû¿ë °¡´ÉÇÑ transduction enhancer:
RetroNectin
¢ç
(Code T100A)
- Lentivirus ³óÃà, ÀϹݿø½ÉºÐ¸®±â·Î 1½Ã°£¸¸¿¡ 100¹è:
Lenti-X¢â Concentrator (Code 631231 ¿Ü)
- ÀçÇö¼º ÀÖ´Â ½ÇÇèÀ» À§Çؼ´Â titrationÀÌ ÇʼöÀÔ´Ï´Ù.
qPCR¹ý:
Lenti-X¢â qRT-PCR Titration Kit (Code 631235)
ELISA¹ý:
Lenti-X¢â p24 Rapid Titer Kit (Single Wash) (Code 631476)
Lateral flow¹ý Á¤·®, 10ºÐ ¿Ï¼º:
Lenti-X¢â GoStix¢â Plus (Code 631280)
Keyword :
Lentivirus
[°³¿ä] Packaging Single Shots
[FAQ] Lentiviral Gene Delivery
Lentiviral expression vector¸¦ Æ÷ÇÔÇÑ ½Ã½ºÅÛ..
CMV promoter º¤ÅÍ ½Ã½ºÅÛ
Á¶Ç÷¸ð¼¼Æ÷, Áٱ⼼Æ÷ µîÀÇ Lentiviral transdu..
EF1 ¥á promoter º¤ÅÍ ½Ã½ºÅÛ
SARS-CoV-2-pseudotyped lentivirus »ý»ê
Lenti-X¢â SARS-CoV-2 Packaging Mix
ÀÓ»ó Ä¡·áÁ¦ °³¹ßÀ» À§ÇÑ
3¼¼´ë Lentivirus packaging system
´Ù¾çÇÑ Çü±¤´Ü¹éÁú ¸®Æ÷Å͸¦ Æ÷ÇÔÇÑ Lentiviral..
Çü±¤´Ü¹éÁú ¸®Æ÷ÅÍ º¤ÅÍ
Tetracycline Á¸Àç ÇÏ¿¡¼¸¸ ¸ñÀû ´Ü¹éÁúÀ» ¹ß..
Tet-on inducible Expression (Two vectors)
Tetracycline Á¸Àç ÇÏ¿¡¼¸¸ ¸ñÀû ´Ü¹éÁúÀ» ¹ß..
Tet One Inducible Expression (One vector)
Lentivirus¸¦ ÀÌ¿ëÇÑ sgRNA, Cas9 gene deliver..
Lenti-X¢â CRISPR/Cas9 System
Tetracycline Á¸Àç ÇÏ¿¡¼¸¸ ¸ñÀû ´Ü¹éÁúÀ» ¹ß..
(Legacy) Tet-Inducible expression ½Ã½ºÅÛ
Pormoterless vector·Î pomorter È°¼º ¿¬±¸
Reporter assay ½Ã½ºÅÛ (Promoterless vector)
Transfection ½Ã¾àÀ» Æ÷ÇÔÇÏ¿© ´õ °£´ÜÇÑ packa..
Packaging single shots
239T, HEK-293 Cellº¸´Ù ´õ ³ôÀº packaging È¿..
Packaging Cell Lines
1 step qRT-PCR·Î ¿ª°¡¸¦ ºü¸£°í Á¤È®ÇÏ°Ô ÃøÁ¤
Lenti-X¢â qRT-PCR Titration Kit
p24 ELISA·Î ºü¸£°í Á¤È®ÇÏ°Ô ¿ª°¡ ÃøÁ¤
Lenti-X¢â p24 Rapid Titer Kit (Single Wash)
½º¸¶Æ®ÆùÀ¸·Î 10ºÐ¸¸¿¡ Lentivirus ¿ª°¡(Titer)..
GoStix¢â Plus
Lentivirus ³óµµ°¡ ³·À» ¶§, Mix & SpinÀ¸·Î °£..
Lentivirus ³óÃà ½Ã¾à
In vivo Lentiviral delivery Àû¿ë ½Ã, ÇʼöÀû..
Lentivirus Á¤Á¦
Ecotropic lentivirus¸¦ human cell¿¡ Àû¿ë
Ecotropic Receptor Booster
Retrovirus ¹× LentivirusÀÇ transduction enha..
RetroNectin¢ç (Recombinant Human Fibronectin Fragment)
Lentivirus³ª MMLV retrovirusÀÇ ´õ ºü¸¥ trans..
Transduction ½Ã¾à
¸ñÀû ¼¼Æ÷ÀÇ À¯ÀüÀÚ¿¡ »ðÀÔµÈ Provirus copy nu..
Provirus Quantitation Kit
Lentivirus°¡ ¸ñÀû¼¼Æ÷ À¯ÀüÀÚ¿¡ »ðÀÔµÈ À§Ä¡¸¦..
Integration Site Analysis Kit
½Å°³³ä lentivirus transduction
Lenti-X¢â Transduction Sponge
ÃÖÀûÈµÈ Transfection Reagent & Packaging Mi..
Trans
IT¢ç Lentivirus System