Cy5 Cas9 mRNA with N1-Me-pUTP (5'CAP) is produced through in vitro transcription. By simulating the mRNA processing process in eukaryotes, Cy5 Cas9 mRNA with N1-Me-pUTP (5'CAP) has a 5 'end Cap 1 cap structure, a 3' end poly (A) tail, Cy5-UTP modification, and N1-Me-pUTP modification (Cy5-UTP: N1-Me-pUTP=3:1 (molar ratio)), which increases the stability and translation efficiency of mRNA[1]. N1-Me-pUTP is a methyl modification of naturally occurring pseudouridine pUTP, catalyzed by N1 specific pseudouridine methyltransferase Nepl present in archaea and eukaryotes[2]. This product uses N1-Me-pUTP instead of UTP, effectively enhancing RNA stability while reducing anti RNA immune response[3]. Cy5 is a commonly used cyanine fluorescent dye with maximum excitation/emission wavelengths of 650/670nm, capable of real-time monitoring of the transfection, localization, and expression of target proteins in cells.
Bacteria and archaea use the CRISPR/Cas system to defend against invading viruses and plasmids. The CRISPR/Cas system functions in eukaryotic systems through two molecular components: Cas9 protein and non coding guide RNA (gRNA). The Cas9 endonuclease reaches the target site under the guidance of gRNA and cleaves the sequence, resulting in gene silencing.
This product should be used in combination with purified Guide RNA.
References:
[1]. Jemielity J, Fowler T, Zuberek J, et al. Novel "anti-reverse" cap analogs with superior translational properties. RNA. 2003;9(9):1108-1122.
[2]. Callum J C Parr, et al.N 1-Methylpseudouridine substitution enhances the performance of synthetic mRNA switches in cells. 2020 Apr 6;48(6):e35. doi: 10.1093/nar/gkaa070.
[3]. Pedro Morais, Hironori Adachi, Yi-Tao Yu.The Critical Contribution of Pseudouridine to mRNA COVID-19 Vaccines. 2021 Nov 4;9:789427. doi: 10.3389/fcell.2021.789427.