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CRISPR/Cas9 Genome Editing: mRNA + gRNA Transfection

TransIT®-mRNA Transfection Kit for mRNA and Guide RNA Oligonucleotide Delivery

Genome Editing

  • In order to avoid off-target cleavage and unwanted genomic integration of plasmid DNA, Cas9-encoding mRNA can be co-transfected with guide RNA oligonucleotides.

Benefits of RNA-based genome editing include:

  • High Specificity – Rapid gene expression generates a transient pulse of genome editing activity
  • Ease of Use – Deliver mRNA and guide RNA with a single reagent
  • DNA-free – No risk of insertional mutagenesis

CRISPR/Cas9 Delivery Methods - Cas9 mRNA + Guide RNA Oligonucleotides

CRISPR mRNA and gRNA Delivery Approaches

CRISPR/Cas9 Delivery Methods – Cas9 mRNA + Guide RNA Oligonucleotides.
Cas9 is supplied as messenger RNA, and guide RNAs are supplied as either synthetic or in vitro transcribed RNA oligonucleotides. (A) The wild-type Cas9 enzyme contains two endonuclease domains which cleave the target DNA on both strands when programmed with a guide RNA. (B) The D10A mutation converts Cas9 into a nickase that generates single-stranded breaks in the target DNA. For improved target specificity, Cas9 D10A can be used with paired guide RNAs targeting opposite strands to create staggered double-stranded breaks.

Efficient Genome Editing with Cas9 mRNA + Guide RNA Oligonucleotides

CRISPR mRNA and gRNA Delivery Data

Efficient Genome Editing with Cas9 mRNA + Guide RNA Oligonucleotides.
HEK293T/17, U2OS, and NHDF cells were co-transfected with 0.5 µg of Cas9 encoding mRNA, 5meC, ψ (Trilink Biotechnologies), and 25 nM of PPIB targeting 2-part gRNA (Dharmacon) using TransIT®-mRNA Transfection Kit (0.5 µl/well of 24-well plate of both mRNA Reagent and Boost, Mirus Bio). A T7E1 mismatch detection assay was used to measure cleavage efficiency at 48 hours post-transfection.

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
MIR2225 TransIT®-mRNA Transfection Kit 0.4 ml 362,00 PDF
MIR2250 TransIT®-mRNA Transfection Kit 1 ml 647,00 PDF
MIR2255 TransIT®-mRNA Transfection Kit 5 x 1 ml 2834,00 PDF
MIR2256 TransIT®-mRNA Transfection Kit 10 x 1 ml 5243,00 PDF
  • All prices are in EURO excl. VAT and shipping. Only available in selected countries.

  • * Sample Policy:
    MoBiTec together with Mirus considers sample requests for up to three different transfection products and one electroporation product per laboratory. Please include all samples to be tested in a single request.

Download

  • CRISPR-Cas9-Poster
  • Mirus CRISPR/Cas9 Transfection Poster (PDF):
    Optimization of DNA, RNA and RNP delivery methods for efficient CRISPR/Cas9 Mediated Mammalian Cell Engineering