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Genome Editing
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ÆíÁý À¯ÀüÀÚ µµÀÔ
> Lenti-X¢â CRISPR/Cas9 System
Lentivirus¸¦ ÀÌ¿ëÇÑ sgRNA, Cas9 gene delivery
Lenti-X¢â CRISPR/Cas9 System
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(ºÎ°¡¼¼º°µµ)
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Clontech
632629
Lenti-X¢â CRISPR/Cas9 System
Each
(Package)
2,211,000¿ø
Clontech
632630
pLVX-hyg-sgRNA1 Vector System
10 Rxns
(Package)
592,000¿ø
Clontech
632633
Lenti-X¢â Tet-On¢ç 3G CRISPR/Cas9 System
1 System
(Package)
3,562,000¿ø
Product index
Overivew
CRISPR/Cas9 Overview
CRISPR ½ÇÇè °¡À̵å (Tu..
About Guide-it¢â Kits
CRISPR Selection Guide
[Cancer Research] Gene..
Customer data for Guid..
Guide-it¢â À¯ÀüÀÚ ÆíÁý..
Knock-in FAQ
À¯ÀüÀÚ ÆíÁý Àü ´Ü°è
sgRNA ÇÕ¼º°ú ½ºÅ©¸®´×
[Upgrade] Guide-it¢â L..
Genome-Wide sgRNA Libr..
ÆíÁý À¯ÀüÀÚ µµÀÔ
CRISPR Plasmid System
CRISPR/Cas9 Gesicle Pr..
AAV CRISPR/Cas9 Vector..
AAV CRISPR/SaCas9 Help..
Lenti-X¢â CRISPR/Cas9 ..
Cas9 Ç×ü (Poly/Mono)
RNA Transfection Reage..
iPSC Single-Cell Cloni..
Recombinant Cas9 ´Ü¹é..
À¯ÀüÀÚ ÆíÁý ÈÄ È®ÀÎ
Mutation Detection Kit
¿°±âġȯ, »ðÀÔ º¯ÀÌ °Ë..
Genotype Confirmation ..
Indel Identification K..
CRE recombinase
CRE Recombinase Gesicl..
AAV Helper Free System..
Cas9 gene°ú sgRNA¸¦ Lentivirus¸¦ ÀÌ¿ëÇÏ¿© ¼¼Æ÷ ³» µµÀÔ
TransfectionÀÌ ¾î·Á¿î mammalian ¼¼Æ÷ (proliferating and non-proliferating cells)¿¡¼ÀÇ CRISPR/Cas9 Genome editing
Lenti-X¢â Packaging Single shotsÀ» ÀÌ¿ëÇÏ¿© °£´ÜÇÏ°Ô °í¿ª°¡ÀÇ Lentivirus Á¦ÀÛ
Lenti-X¢â Tet-On 3G CRISPR/Cas9 systemÀº Cas9 ¹ßÇöÀ» È¿À²ÀûÀ¸·Î Á¶ÀýÀÌ °¡´ÉÇϸç, ¼¼Æ÷ µ¶¼º ¹®Á¦³ª Cas9ÀÇ Áö¼ÓÀûÀÎ ¹ßÇöÀ¸·Î ÀÎÇÑ off-target effect °¨¼Ò¿¡ È¿°úÀû
Á¦Ç°¼³¸í
Lenti-X¢â CRISPR/Cas9 SystemÀº lentiviral-mediated CRISPR/Cas9 genome editingÀ» À§ÇÑ ¸ðµç ±¸¼ºÇ°ÀÌ Æ÷ÇÔµÈ complete systemÀÌ´Ù.
º» Á¦Ç°Àº °í¿ª°¡ÀÇ ·»Æ¼¹ÙÀÌ·¯½º¸¦ »ý»êÇÏ´Â Lenti-X¢â Packaging Single Shots°ú Cas9 ´Ü¹éÁú, sgRNA¸¦ ¹ßÇöÇÏ´Â µÎ °³ÀÇ Çö󽺹̵å·Î ±¸¼ºµÇ¾îÀÖ´Ù. ¸ñÀû À¯ÀüÀÚ¸¦ Ÿ°ÙÇÏ´Â sgRNA¸¦ ¹ßÇöÇÏ´Â pLVX-hgy-sgRNA1 plasmid´Â ÀÌ¹Ì ¼±ÇüÈ µÇ¾îÀÖ¾î ¾ÆÁÖ ½±°Ô sgRNA ¼¿À» »ðÀÔ, ¶óÀÌ°ÔÀ̼ÇÀ» ÁøÇàÇÒ ¼ö ÀÖ´Ù. Lentivirus¸¦ ÀÌ¿ëÇÏ¿© sgRNA¿Í Cas9 geneÀ» »ðÀÔÇϸé ÀϹÝÀûÀ¸·Î TransfectionÀÌ ¾î·Æ´Ù°í ¾Ë·ÁÁø ¼¼Æ÷ÁÖ¿¡¼ÀÇ CRISPR/Cas9 genome editingÀÌ °¡´ÉÇÏ´Ù (±×¸² 1).
Lenti-X¢â Tet-On 3G CRISPR/Cas9 SystemÀº doxycyclineÀ» ÀÌ¿ëÇÏ¿© Cas9 ¹ßÇöÀ» À¯µµÇÒ ¼ö ÀÖ´Â Tet-on 3G inducible expression Á¦Ç°À¸·Î½á, doxycycline ó¸® ½Ã Cas9 ´Ü¹éÁúÀ» ¹ßÇöÇÑ´Ù. Cas9 ¹ßÇöÀ» ¾ö°ÝÇÏ°Ô Á¶ÀýÇÒ ¼ö Àֱ⠶§¹®¿¡ ¿µ±¸ÀûÀÎ Cas9 ´Ü¹éÁú ¹ßÇöÀ¸·Î ÀÎÇÑ off-target effect³ª ¼¼Æ÷ µ¶¼ºÀ» ÁÙÀÏ ¼ö ÀÖ´Ù (±×¸² 2, ±×¸² 3).
±×¸² 1. Either Jurkat or HT1080 cells were first transduced with LVX-hyg-CD81-sgRNA and then selected for stable integration using hygromycin.
Stable clones were then transduced with LVX-puro-Cas9. Stable clones were selected for using puromycin and then screened for CD81 knockout efficiency using FACS. Positive and negative controls of parental cells without Cas9 transduction were done either with or without anti-CD81 antibody.
±×¸² 2. Inducible knockout of
CD81
in HEK293 cells.
Target cells were first transduced with Tet-On 3G lentivirus and selected with G418. Cas9 induction was then assayed by Western blot using
polyclonal anti-Cas9 antibody (Code 632607)
, diluted 1:1,000, and ECL reagent.
Panel A.
Western blot results for six independent Tet-On 3G-Cas9-positive cell populations cultured in the absence (-) or presence (+) of 0.5 ¥ìg/ml doxycycline, assayed with anti-Cas9 antibody.
Panel B.
FACS results for three independent Tet-On 3G-Cas9-positive 293T cell populations transduced with CD81-sgRNA lentivirus and cultured in the absence (-Dox) or presence (+Dox) of 0.5 ¥ìg/ml doxycycline.
±×¸² 3. Inducible editing of
AAVS1
in HEK293 cells.
Induction of Cas9 expression in six stable Tet-On 3G-Cas9-positive HEK293 cell lines was assayed by qRT-PCR. The
Guide-it Mutation Detection Kit (Code 631443)
was then used to screen for editing of
AAVS1
.
Panel A.
qRT-PCR results for Cas9 expression in uninduced (-Dox) and induced (+Dox) cells for each clone.
Panel B.
Results of Guide-it resolvase assay for detection of
AAVS1
gene editing in uninduced (?) and induced (+) clones.
Applications
Lentivirus-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)
Lenti-X CRISPR/Cas9 System (Code 632629)
pLVX-hyg-sgRNA1 Vector (Linear) (
Vector map
/
Full sequence (txt.)
)
pLVX-puro-Cas9 Vector (
Vector map
/
Full sequence (txt.)
)
Stellar Competent Cells
Guide-it Ligation Components v2
Lenti-X Packaging Single Shots
pLVX-hyg-sgRNA1 Vector System (Code 632630)
pLVX-hyg-sgRNA1 Vector (
Vector map
/
Full sequence (txt.)
)
Guide-it Ligation Components v2
Stellar Competent Cells
Lenti-X
¢â Tet-On 3G CRISPR/Cas9 System (Code 632633)
pLVX-hyg-sgRNA1 Vector (Linear) (
Vector map
/
Full sequence (txt.)
)
pLVX-TRE3G-Cas9-puro Vector Set
- pLVX-TRE3G-Cas9-puro Vector (
Vector map
/
Full sequence (txt.)
)
- pLVX-TRE3G-Luc Control Vector (
Vector map
/
Full sequence (txt.)
)
pLVX-EF1a-Tet3G Vector (
Vector map
/
Full sequence (txt.)
)
Stellar Competent Cells
Guide-it Ligation Components v2
Lenti-X Packaging Single Shots
Keyword : Lenti,Lentivirus,Lentiviral,Tet,Tetracycline,inducible,inducible system,µµÀÔÈ¿À²,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