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Home > 전제품보기 > Genome Editing > CRISPR-Cas9 gene delivery > Xfect™ RNA Transfection Reagent
mRNA, sgRNA, siRNA등 다양한 RNA transfection에 최적

Xfect™ RNA Transfection Reagent

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제조사 제품코드 제품명 용량 가격
(부가세별도)
비고 사용자매뉴얼
Clontech
631450
Xfect™ RNA Transfection Reagent
관련학술 관심상품등록 구매하기 라이선스 
1.2 ml
400,000원  제조사 페이지로 바로가기 109673

  • CRISPR/Cas9을 위한 sgRNA 및 mRNA, miRNA, siRNA 등 여러 종류의 RNA Transfection에 최적
  • 다양한 종류의 포유류 세포 (mammalian cell)에 적용 가능
  • 높은 효율의 유전자 도입과 매우 낮은 세포 독성
  • Simple, serum-compatible protocol
  • Transfection에 필요한 모든 구성품을 포함한 All-in-One system
제품설명
Xfect™ RNA Transfection reagent는 다양한 종류의 mammalian cell (broad-range)에 microRNA, siRNA, mRNA 또는 sgRNA와 같은 여러 종류의 RNA를 효율적으로 도입 가능한 Transfection 시약이다. 생분해성 나노입자 (biodegradable nanoparticles)로 구성된 Xfect™ Transfection reagnet는 일반적인 Transfection 시약에 비하여 낮은 세포독성 및 높은 도입효율로 RNA Transfection이 가능하다.
특히 본 제품은 CRISPR/Cas9-mediated gene editing에 사용되는 Cas9 coding mRNA 및 single guide RNA 도입에 효율적으로 사용이 가능하며, Xfect™ Transfection reagent (Code 631317, 631318)과 함께 사용하면, DNA와 RNA co-transfection이 가능하다.



그림 1. Xfect RNA Transfection 시약을 이용한 sgRNA Transfection 후, CD81 gene knock-out 결과
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 105 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.



그림 2. Primary cell 및 다양한 종류의 cell line에서의 mRNA transfection 효과
HeLa cells (2.0 x 105), HEK 293 cells (1.5 x 105), Human Dermal Fibroblasts (NHDF) cells (6.0 x 104), Mesenchymal Stem Cells (MSCs) (5.0 x 104), Jurkat Cells (3.0 x 105), and KBM7 cells (3.0 x 105) were plated in 12-well plates and transfected with 1 μg of mRNA encoding GFP with 5 μl 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.



그림 3. Primary cell 및 cell line에서 siRNA Transfection에 의한 Knock-down 효율
HeLa cells (2.0 x 105), Human Dermal Fibroblasts (NHDF) cells (6.0 x 104), and Mesenchymal Stem Cells (MSCs) (5.0 x 104) 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 μg 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
구성품 (자세한 내용은 CoA를 참조하세요)

Xfect™ RNA Transfection Polymer

600ul × 2

Xfect™ Reaction Buffer

12ml × 2

보존 - 20 ℃

Keyword : RNA transfection,트랜스펙션,CRISRP/Cas9,genome editing,sgRNA,Xfect
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