Cas9 mRNA with N1-Me-pUTP (5'CAP) is produced through in vitro transcription. By simulating the mRNA processing process in eukaryotes, it has a 5' end Cap 1 cap structure and a 3' end poly(A) tail, which increases the stability and translation efficiency of the mRNA[1]. N1-methyl-pseudouridine (1-methylpseudouridine, m1ψ) is a methyl modification of naturally occurring pseudouridine, formed by N1-specific pseudouridine A that exists in archaea and eukaryotes It is catalyzed by the base transferase Nepl[2]. N1-methyl-pseudouridine is a substitute for uridine (U), which can effectively enhance RNA stability while reducing anti-RNA immune response [3].
Bacteria and archaea use the CRISPR/Cas system to defend against invading viruses and plasmids. The CRISPR/Cas system works in eukaryotic systems through two molecular components: Cas9 protein and non-coding guide RNA (gRNA). Cas9 endonuclease reaches the target site under the guidance of gRNA and cuts the sequence, resulting in gene silencing.
This product should be used in conjunction 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.
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,该产品具有5'端Cap 1帽结构和3'端poly(A)尾,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]。该产品使用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]。
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。