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Genome Editing
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ÆíÁý À¯ÀüÀÚ µµÀÔ
> RNA Transfection Reagent
mRNA, sgRNA, siRNAµî ´Ù¾çÇÑ RNA transfection¿¡ ÃÖÀû
RNA Transfection Reagent
Á¦Á¶»ç
Á¦Ç°ÄÚµå
Á¦Ç°¸í
¿ë·®
°¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í
»ç¿ëÀڸŴº¾ó
Clontech
631450
Xfect¢â RNA Transfection Reagent
1.2 ml
528,000¿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..
CRISPR/Cas9À» À§ÇÑ sgRNA ¹× mRNA, miRNA, siRNA µî ¿©·¯ Á¾·ùÀÇ RNA Transfection¿¡ ÃÖÀû
´Ù¾çÇÑ Á¾·ùÀÇ Æ÷À¯·ù ¼¼Æ÷ (mammalian cell)¿¡ Àû¿ë °¡´É
³ôÀº È¿À²ÀÇ À¯ÀüÀÚ µµÀÔ°ú ¸Å¿ì ³·Àº ¼¼Æ÷ µ¶¼º
Simple, serum-compatible protocol
Transfection¿¡ ÇÊ¿äÇÑ ¸ðµç ±¸¼ºÇ°À» Æ÷ÇÔÇÑ All-in-One system
Á¦Ç°¼³¸í
Xfect¢â RNA Transfection reagent´Â ´Ù¾çÇÑ Á¾·ùÀÇ mammalian cell (broad-range)¿¡ microRNA, siRNA, mRNA ¶Ç´Â sgRNA¿Í °°Àº ¿©·¯ Á¾·ùÀÇ RNA¸¦ È¿À²ÀûÀ¸·Î µµÀÔ °¡´ÉÇÑ Transfection ½Ã¾àÀÌ´Ù. »ýºÐÇؼº ³ª³ëÀÔÀÚ (biodegradable nanoparticles)·Î ±¸¼ºµÈ Xfect¢â Transfection reagnet´Â ÀϹÝÀûÀÎ Transfection ½Ã¾à¿¡ ºñÇÏ¿© ³·Àº ¼¼Æ÷µ¶¼º ¹× ³ôÀº µµÀÔÈ¿À²·Î RNA TransfectionÀÌ °¡´ÉÇÏ´Ù.
ƯÈ÷ º» Á¦Ç°Àº CRISPR/Cas9-mediated gene editing¿¡ »ç¿ëµÇ´Â Cas9 coding mRNA ¹× single guide RNA µµÀÔ¿¡ È¿À²ÀûÀ¸·Î »ç¿ëÀÌ °¡´ÉÇϸç,
Xfect¢â Transfection reagent (Code 631317, 631318)
°ú ÇÔ²² »ç¿ëÇϸé, DNA¿Í RNA co-transfectionÀÌ °¡´ÉÇÏ´Ù.
±×¸² 1. Xfect
¢â
RNA Transfection
½Ã¾àÀ» ÀÌ¿ëÇÑ sgRNA Transfection ÈÄ, CD81 gene knock-out °á°ú
Panel A.
sgRNA targeting the 5¡¯ end of the antisense strand of
CD81
was synthesized using the
Guide-it sgRNA
In Vitro
Transcription Kit
.
Panel B.
An HT1080 cell line (2.0 x 10
5
cells) stably expressing Cas9 (HT1080-Cas9) was transfected with 50 pmol of sgRNA targeting
CD81
, either once or twice (lower graph), using the Xfect RNA Transfection Reagent. Seven days later, cells were immunostained with a CD81 antibody (Ab) conjugated to an FITC fluorophore and analyzed by flow cytometry. The percentage of cells that did not bind CD81 was calculated. A control sample, comprised of HT1080-Cas9 cells, was analyzed by flow cytometry, either without (top, left graph) or with (top, right graph) the CD81 antibody. Both single and double transfection with sgRNA resulted in a substantial increase in cells that did not bind CD81, indicating successful CRISPR/Cas9-mediated knockout of
CD81
.
±×¸² 2. Primary cell ¹× ´Ù¾çÇÑ Á¾·ùÀÇ cell line¿¡¼ÀÇ mRNA transfection È¿°ú
HeLa cells (2.0 x 10
5
), HEK 293 cells (1.5 x 10
5
), Human Dermal Fibroblasts (NHDF) cells (6.0 x 10
4
), Mesenchymal Stem Cells (MSCs) (5.0 x 10
4
), Jurkat Cells (3.0 x 10
5
), and KBM7 cells (3.0 x 10
5
) were plated in 12-well plates and transfected with 1 ¥ìg of mRNA encoding GFP with 5 ¥ìl of Xfect RNA Transfection Reagent. 20 hours later, cells were analyzed by flow cytometry and the % GFP-positive cells and the Mean Fluorescence Intensity (MFI) were determined.
±×¸² 3. Primary cell ¹× cell line¿¡¼ siRNA Transfection¿¡ ÀÇÇÑ Knock-down È¿À²
HeLa cells (2.0 x 10
5
), Human Dermal Fibroblasts (NHDF) cells (6.0 x 10
4
), and Mesenchymal Stem Cells (MSCs) (5.0 x 10
4
) were plated in 12-well plates and transfected with 50 pmol of siRNA against
luciferase
using Xfect RNA Transfection Reagent. All three cell types were also transfected with 1 ¥ìg of pCMV-Luc using the
Xfect Transfection Reagent
. Luciferase assays were performed 48 hours post-transfection. For control samples, cells were transfected with pCMV-Luc but without the siRNA. We observed a dramatic (>95%) decrease in luciferase activity in all the cells treated with siRNA.
Applications
CRISPR/Cas9
RNA transfection
Transient transfection without genomic integration
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
Xfect¢â RNA Transfection Polymer
600ul ¡¿ 2
Xfect¢â Reaction Buffer
12ml ¡¿ 2
º¸Á¸ - 20 ¡É
Keyword : RNA transfection,Cas9 RNA,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