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mRNA, sgRNA, siRNAµî ´Ù¾çÇÑ RNA transfection¿¡ ÃÖÀû

RNA Transfection Reagent

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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
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1.2 ml
528,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33197

  • 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 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.



±×¸² 2. Primary cell ¹× ´Ù¾çÇÑ Á¾·ùÀÇ cell line¿¡¼­ÀÇ mRNA transfection È¿°ú
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 ¥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 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 ¥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

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