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.
Cy5 Cas9 mRNA with N1-Me-pUTP (5'CAP)是通过体外转录产生的，通过模拟真核生物中mRNA加工过程具有5'端Cap 1帽结构、3'端poly(A)尾、Cy5-UTP修饰以及N1-Me-pUTP修饰（Cy5-UTP：N1-Me-pUTP=3:1（摩尔比）），增加了mRNA的稳定性和翻译效率[1]。N1-Me-pUTP是天然存在的假尿苷pUTP的甲基修饰物，由存在于古细菌和真核生物中的N1特异性假尿苷甲基转移酶Nepl催化生成[2]。本产品使用N1-Me-pUTP替代UTP，有效增强了RNA稳定性，同时降低抗RNA免疫应答[3]。Cy5是一种常用的花青类荧光染料，最大激发/发射光波长分别为650/670nm，能够实时监测细胞中目的蛋白的转染、定位和表达情况。
细菌和古菌使用CRISPR / Cas系统来防御入侵的病毒和质粒。CRISPR/Cas系统通过两种分子成分在真核系统中发挥作用：Cas9蛋白和非编码向导RNA(Guide RNA,gRNA)。Cas9核酸内切酶在gRNA的导向下到达目的位点并切割序列，从而导致基因沉默。
本产品应与纯化的Guide RNA结合使用。