´Ý±â
TaKaRa
Á¦Ç°¸í/Á¦Ç°ÄÚµå
Å°¿öµå°Ë»ö
°Ë»ö
´Ý±â
  • °í°´Áö¿ø 
  • ¾÷¹«¾È³»¦¢Á¦Ç°¹®ÀÇ
  • ÀüÈ­¹øÈ£¦¢02-2081-2510
  • Email     ¦¢support@takara.co.kr
  • ´ëÀüÁö»ç 
  • ¾÷¹«¾È³»¦¢´ëÀü/ÃæûÁö¿ª ÁÖ¹®, Á¦Ç°¹®ÀÇ
  • ÀüÈ­¹øÈ£¦¢042-828-6525
  • Email     ¦¢tkbd@takara.co.kr
  • ¾÷¹«½Ã°£¾È³»
  • [ Æò¡¡¡¡ÀÏ ] 09 : 00 ~ 18 : 00 ¦¢ [ Á¡½É½Ã°£ ] 12 : 00 ~ 13 : 00
  • Å䡤ÀÏ¿äÀÏ, °øÈÞÀÏÀº ÈÞ¹«ÀÔ´Ï´Ù.
Single vector¸¦ ÀÌ¿ëÇÑ È¿À²ÀûÀÎ sgRNA, Cas9 ¹ßÇö

CRISPR Plasmid System

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
632601
Guide-it¢â CRISPR/Cas9 System (Green)
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç°  ¶óÀ̼±½º 
1 System
(Package)
728,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32687, x33376
Clontech
632602
Guide-it¢â CRISPR/Cas9 System (Red)
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç°  ¶óÀ̼±½º 
1 System
(Package)
728,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32687, x33375

  • Single vector system: ÇϳªÀÇ º¤ÅÍ¿¡¼­ target-specific sgRNA, Cas9 nuclease, Çü±¤´Ü¹éÁú (ZsGreen1 ¶Ç´Â tdTomato) ¹ßÇö
  • Exceptionally bright fluorescent protein: trasnfection È¿À²À» È®ÀÎÇÒ ¼ö ÀÖ´Â ¸®Æ÷Åͷνá EGFPº¸´Ù 2.5¹è~6¹è ´õ ¹àÀº ZsGreen1
    ¶Ç´Â tdTomato »ç¿ë
  • Simple Plasmid construction: vector°¡ ¹Ì¸® ¼±ÇüÈ­(Pre-linearized vector)µÇ¾îÀÖ¾î ¿øÇÏ´Â À¯ÀüÀÚÀÇ ÇÕ¼º sgRNA¸¦ °£´ÜÈ÷
    ligation & clone
  • Complete system: sgRNA/Cas9 system¿¡ ÇÊ¿äÇÑ Á¦Ç°ÀÌ ¸ðµÎ Æ÷ÇÔ

Á¦Ç°¼³¸í
Guide-it CRISPR/Cas9 system Á¦Ç°Àº CRISPR/Cas9 ±â¼úÀÇ mammalian genome modification (or genome editing)À» À§ÇØ target single guide RNAs(sgRNAs)ÀÇ Å¬·Î´× ¹× ¹ßÇöÀ» ÇÒ ¼ö ÀÖµµ·Ï Á¦ÀÛµÈ Á¦Ç°ÀÌ´Ù. º» Á¦Ç°¿¡ Æ÷ÇԵǾîÀÖ´Â º¤ÅÍ´Â Cas9 nuclease, target-specific sgRNA, Çü±¤´Ü¹éÁúÀ» ¹ßÇöÇÒ ¼ö ÀÖÀ¸¸ç, ¹ßÇöµÈ Çü±¤´Ü¹éÁúÀº transfection È¿À²À» ¸ð´ÏÅ͸µ Çϰųª Flow cytometry¸¦ ÀÌ¿ëÇÏ¿© transfected cellÀ» isolating/enriching ¿¡ »ç¿ëÇÒ ¼ö ÀÖ´Ù.



±×¸² 1. The structure of the pGuide-it vector. This vector is part of the Guide-it CRISPR/Cas9 Systems, kits for the cloning of a target single guide RNA (sgRNA) for CRISPR/Cas9 genome editing. The pGuide-it vector is used to simultaneously express Cas9 nuclease, a target-specific sgRNA, and an exceptionally bright fluorescent protein (either ZsGreen1 or tdTomato). Cas9 and the fluorescent protein are co-expressed from a bidirectional CMV promoter, and a user-defined sgRNA is cloned downstream of a human U6 promoter. To construct the vector, a pair of oligos corresponding to the genomic target (your guide sequence) are annealed to form a duplex. Then, the duplexed DNA is cloned into the pre-linearized vector using the included high-efficiency ligation mix and competent cells.



±×¸² 2. Genomic DNA modifications introduced using the pGuide-it-ZsGreen1 plasmid. 1 x 105 cells (HEK 293, HeLa, U2OS, or HT1080 cell lines) were seeded in 12-well plates 16 hr prior to transfection. Cells were transfected with 2.5 ¥ìg of pGuide-it-ZsGreen1 plasmid using the Xfect Transfection Reagent (Code 631317); the plasmid expressed Cas9, ZsGreen1, and an sgRNA targeting the either CCR5, C4BPB, or EMX1. After 48 hr, fluorescence was analyzed by microscopy (A), and by FACS (B, Transfection efficiency). The efficiency of gene modification was also evaluated using the Guide-it Mutation Detection Kit (Code 631443), a PCR-based method for testing for the presence of indels (B). The target sequence was amplified directly from cells, the amplicon was melted and hybridized to form mismatched targets that were cleaved with Guide-it Resolvase. Cleavage products were resolved on an agarose gel and quantified by densitometry to estimate the frequency of indels (B, Indel).
Àû¿ë
  • Cloning and expression of a target single guide RNA (sgRNA) for mammalian genome editing using CRISPR/Cas9 technology
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
Guide-it CRISPR/Cas9 System (Green) (Cat. No. 632601)
  • pGuide-it-ZsGreen1 Vector (Linear)
  • Guide-it Ligation Components
  • Stellar Competent Cells

Guide-it CRISPR/Cas9 System (Red) (Cat. No. 632602)
  • pGuide-it-tdTomato Vector (Linear)
  • Guide-it Ligation Components
  • Stellar Competent Cells
º¸Á¸
Stellar Competent Cells:-70¡É
±âŸ:-20¡É




Keyword : CRISPR vector,Cas9 vector,CRISPR,CRISPR/Cas9,Cas9,sgRNA,Å©¸®½ºÆÛ,gene editing,À¯ÀüÀÚÆíÁý

ÆíÁý À¯ÀüÀÚ µµÀÔ
>
Single vector¸¦ ÀÌ¿ëÇÑ È¿À²ÀûÀÎ sgRNA, Cas9 ..
CRISPR Plasmid System
>
Off-target effect °¨¼Ò, Hard-to-transfect ce..
CRISPR/Cas9 Gesicle Production System
>
Adeno-associated virus delivery-CRISRP/Cas9 ..
AAV CRISPR/Cas9 Vector System
>
SaCas9¸¦ ÀÌ¿ëÇÑ AAV mediated CRISPR Genome E..
AAV CRISPR/SaCas9 Helper Free System (AAV2)
>
Lentivirus¸¦ ÀÌ¿ëÇÑ sgRNA, Cas9 gene deliver..
Lenti-X¢â CRISPR/Cas9 System
>
Cas9 nuclease °ËÃâÀ» À§ÇÑ Ç×ü
Cas9 Ç×ü (Poly/Mono)
>
mRNA, sgRNA, siRNAµî ´Ù¾çÇÑ RNA transfection..
RNA Transfection Reagent
>
hiPS cellÀÇ single cell ¹è¾ç, À¯ÀüÀÚ ÆíÁý¿¡ ..
iPSC Single-Cell Cloning DEF-CS¢â Culture Media Kit
>
Electroporation delivery¸¦ À§ÇÑ Cas9 ´Ü¹éÁú
Recombinant Cas9 ´Ü¹éÁú