¿À´Ã ÇÏ·ç ±×¸¸º¸±â
´Ý±â
·Î±×ÀÎ
|
ȸ¿ø°¡ÀÔ
|
ID/PW ã±â
|
Á¦Ç°»ó´ã¹®ÀÇ
°í°´Áö¿ø
¾÷¹«¾È³»¦¢Á¦Ç°¹®ÀÇ
ÀüȹøÈ£¦¢
02-2081-2510
Email ¦¢
support@takara.co.kr
´ëÀüÁö»ç
¾÷¹«¾È³»¦¢´ëÀü/ÃæûÁö¿ª ÁÖ¹®, Á¦Ç°¹®ÀÇ
ÀüȹøÈ£¦¢
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.
Á¦Ç°Á¤º¸
°øÁö
±â¼úÁö¿ø
Á¤º¸ÀÚ·á
Learning Center
ȸ»çÁ¤º¸
ÀüÁ¦Ç°º¸±â
Á¦Ç°¼±Åð¡À̵å
Custom Service
À̺¥Æ®/ÇÁ·Î¸ð¼Ç
´ÙÄ«¶ó¼Ò½Ä
Takara Bio°ø½Ã
½ÅÁ¦Ç°¾È³»
¶óÀ̼±½º¾È³»
±â¼úÁö¿ø¹®ÀÇ
±â±âµ¥¸ð½Åû
¼¼¹Ì³ª½Åû
LMO¾È³»
ºê·Î¼Å
On-line tool
eNews
樨毢
CoA & MSDS
°³¿ä & ¸®½ºÆ®
¼Ò°³
µµÀԺ귣µå Á¤º¸
´ë¸®Á¡Á¤º¸
ã¾Æ¿À½Ã´Â±æ
Home
>
ÀüÁ¦Ç°º¸±â
>
Gene transfer (virus)
>
Adeno-Associated Virus (AAV)
> CRISPR/Cas9 Vector System
Adeno-associated virus delivery-CRISRP/Cas9 system (Two vector)
CRISPR/Cas9 Vector System
Á¦Á¶»ç
Á¦Ç°ÄÚµå
Á¦Ç°¸í
¿ë·®
°¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í
»ç¿ëÀڸŴº¾ó
Clontech
632608
AAVpro¢ç CRISPR/Cas9 Helper Free System (AAV2)
1 System
(Package)
1,707,000¿ø
,
Clontech
632609
AAVpro¢ç CRISPR/Cas9 Vector System
1 System
(Package)
822,000¿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..
¡Ø SaCas9 (from S. aureus) ¸¦ ÀÌ¿ëÇÑ AAV meditated CRISPR/Cas9 Genome EditingÀº
AAVpro¢ç CRISPR/SaCas9 Helper Free System (AAV2) (Code 632619)
À» È®ÀÎÇϼ¼¿ä.
Adeno-Associated Virus (AAV)¸¦ ÀÌ¿ëÇÑ ¼¼Æ÷ ³» Cas9, sgRNA À¯ÀüÀÚ µµÀÔ
TransfectionÀÌ ¾î·Á¿î Mammalian cell (ºÐ¿, ºñºÐ¿ ¼¼Æ÷)¿¡¼ Genome Editing
AAV¿¡ ÀÇÇÑ Cas9 µµÀÔÀº genome integrationÀ» ¾ïÁ¦ÇÏ°í Cas9ÀÇ ¹ßÇöÀ» Áö¼Ó½Ãų »Ó¸¸ ¾Æ´Ï¶ó, off-target effect¸¦ ¾ïÁ¦
Á¦Ç°¼³¸í
AAVpro¢ç CRISPR/Cas9 Helper free systemÀº CRISPR/Cas9 genome editing¿¡ »ç¿ëµÇ´Â single guide RNA (sgRNA)¿Í Cas9 À¯ÀüÀÚ¸¦ ¹ßÇöÇÏ´Â AAV2 (Adeno-Associated virus)¸¦ Helper virus ¾øÀÌ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. º» Á¦Ç°Àº 4.1kbÀÇ Cas9 À¯ÀüÀÚ¸¦ 2Á¾·ùÀÇ AAV º¤ÅÍ (pAAV-Guide-it-Up, pAAV-Guide-it-Down vector)¿¡ ³ª´©¾î Á¦ÀÛÇÔÀ¸·Î½á AAVÀÇ »çÀÌÁî Á¦ÇÑÀ» ±Øº¹ÇÏ¿´À» »Ó¸¸ ¾Æ´Ï¶ó ´Ù¾çÇÑ mammalin cell in vitro ±¸Ãà ¹× in vivo¿¡¼ÀÇ È¿À²ÀûÀÎ Genome EditingÀ» °¡´ÉÄÉ ÇÏ¿´´Ù. º» Á¦Ç°Àº ÃÑ 10ȸºÐÀÇ sgRNA ¹ßÇöÀ» À§ÇÑ linearized vector, °íÈ¿À²ÀÇ Ligation mix¿Í Stellar competent cellÀÌ Æ÷ÇԵǾîÀÖÀ¸¸ç, modified AAV2 Á¦ÀÛÀ» À§ÇÑ pRC2-mi342 Vector¿Í pHelper vector, AAV2 ÀÔÀÚ ÃßÃâÀ» À§ÇÑ AAV Extraction Solutionµµ Æ÷ÇÔÇÏ°í ÀÖ´Ù.
Figure 1. Production of full-length Cas9 and sgRNA
. The pAAV-Guide-it-Up and pAAV-Guide-it-Down vectors encode truncated upstream and downstream portions of the Cas9 gene respectively, with a 1.6-kb overlapping region of homology.
Figure 2. AAV vector recombination to produce full-length Cas9 and sgRNA
. Separate viruses are generated with each plasmid. Transduction of target cells with both viruses results in recombination to generate the full length Cas9 (4.1 kb) gene. Subsequent expression in target cells results in functional Cas9 protein guided to the appropriate genomic site by the sgRNA.
Figure 3. Vector Map of pAAV-Guide-it-Down.
The structure of the pre-linearized pAAV-Guide-iT-Down vector. This vector is part of the AAVpro CRISPR/Cas9 Helper Free system, a kit that produces AAV particles for cell delivery of the Cas9 and sgRNA genes, components necessary for CRISPR/Cas9 genome editing. The large size of the Cas9 gene precludes its packaging into a single AAV virus. To allow AAV-mediated delivery of the Cas9 gene, this system uses two separate Cas9 plasmids. The pAAV-Guide-it-Down vector contains a truncated downstream portion of the gene (2,616 bp) that encodes the C-terminal 872 amino acids of Cas9. A user-defined single guide RNA (sgRNA) can be cloned into the pAAV-Guide-it-Down vector downstream of the human U6 promoter. To construct the vector, a pair of oligos corresponding to the genomic target (guide sequence) are annealed to form a duplex. The duplexed DNA is then cloned into the pre-linearized vector.
Applications
AAV-based delivery of a user-defined sgRNA and Cas9 for mammalian genome editing using CRISPR/Cas9 technology
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
AAVpro¢ç CRISPR/Cas9 Helper Free System (AAV2) (Cat. No. 632608)
AAVpro¢ç CRISPR/Cas9 Vector Set
Guide-it Ligation Components
Stellar Competent Cells
pRC2-mi342 Vector
pHelper Vector
AAVpro¢ç Extraction Solution
AAVpro¢ç CRISPR/Cas9 Vector System (Cat. No. 632609)
AAVpro¢ç CRISPR/Cas9 Vector Set
Guide-it Ligation Components
Stellar Competent Cells
Keyword :
Adeno-Associated Virus (AAV)
Overview - All about AAV
Adeno-associated virus Á¦ÀÛ¿¡ ÃÖÀûÀÎ packagi..
Packaging (293T Cell Line)
Helper Free ½Ã½ºÅÛ¿¡ ÀÇÇÑ ´Ù¾çÇÑ serotypeÀÇ ..
AAV1, 2, 5, 6 Vector System
Cre-Recombinase À¯ÀüÀÚ¸¦ žÀçÇÑ AAV1, AAV2, ..
CRE Recombinase System
¥â-galactosidase À¯ÀüÀÚ¸¦ žÀçÇÑ AAV1, AAV2,..
LacZ System
Çü±¤´Ü¹éÁúÀÌ Æ÷ÇÔµÈ AAV vector
pAAV-ZsGreen1 Vector
Adeno-associated virus delivery-CRISRP/Cas9 ..
CRISPR/Cas9 Vector System
SaCas9¸¦ ÀÌ¿ëÇÑ AAV mediated CRISPR Genome E..
CRISPR/SaCas9 Vector System (AAV2)
AAV2 Delivery Tet À¯µµ¹ßÇö ½Ã½ºÅÛ
Tet-Inducible System (AAV2)
Cell°ú ¹è¾ç ¹èÁö¿¡¼ AAV ÃßÃâºÎÅÍ Á¤Á¦±îÁö
Cell & Sup. Purification Kit
´Ù¾çÇÑ serotypeÀÇ AAV Á¤Á¦¿¡ ÃÖÀû
Purification Kit Maxi/Midi (All Serotypes)
AAV Serotype 2 Á¤Á¦ kit
Purification Kit (AAV2 Àü¿ë)
AAV¸¦ È¿À²ÀûÀ¸·Î ³óÃà
AAVpro¢ç Concentrator
ºü¸£°í Á¤È®ÇÑ AAV ¿ª°¡ÃøÁ¤ (qPCR)
AAVpro¢ç Titration Kit (for Real Time PCR)
°£ÆíÇÑ AAV ÃßÃâ½Ã¾à
Extraction Solution
±âÁ¸ÀÇ AAV ÃßÃâ ¹æ¹ýÀ¸·Î Á¤Á¦±îÁö 4½Ã°£ ³» ..
AAV Freeze-thaw extraction solution