LZCap™ AG(3'Acm) is a Cap1 analog, can be used as the capping agent for producing mRNAs in an "one-pot"process. Under the action of T7 polymerase, mRNA with 5' end Cap 1 structure was generated by cotranscription using LZCap™ AG(3'Acm), NTPs, and template DNA. The capped mRNA could be directly translated and expressed in cells and in vivo. It is widely used in the fields of in vitro transcription, gene editing, vaccine development, tumor CAR-T therapy, protein substitution therapy and regenerative medicine.
LZCap™ AG(3'Acm) requires the T7 promoter with AG as the starting sequence. LZCap™ AG(3'Acm) can provide>97%capped mRNA, and up to 100-200 μg capped mRNA can be generated with 1 μg DNA template per standard reaction

Specifications

Components

Storage

This product should be stored at -25~-15℃ for 2 years

Figures

With LZCap AG(3'Acm) cap, the luciferase mRNA capping efficiency is about 97.59%, and up to 200 μg capped mRNA can be obtained with 1 μg DNA template in a standard in vitro transcription (IVT) reaction with T7 RNA polymerase. The mRNA purity is up to 99% after simple LiCl precipitation. This high capping efficiency and yield make LZCap an attractive option for various applications, including protein production and gene therapy.

Figure 1. MS analysis of the capping efficiency of LZCapped Luciferase mRNA with RNAse H based method. The capping efficiency is about 97.59%.

Q1. How’s the mRNA stability towards the decapping enzyme

mRNAs with LZCap AG(3'Acm) and LZCap AG M6 (3'Acm) show higher resistance towards the decapping enzyme (NEB). It is noted that CapM6 (3'-OMe-6mA-7mG) is also resistant to decapping enzym, but the regular Cap AG (3'-OMe-7mG) does not show the resistance.

Due to the high affinity of LZCap, the low input of cap1 analog shows perfect performance when used alongside our Hieff™ T7 RNA polymerase (Cat: 10628).

Q2. How’s the binding affinity of LZCap to eIF4E?

Q3. How about the protein expression of LZCap AG(3'Acm) capped mRNA?

The expression of LZCap AG(3'Acm) capped luciferase mRNA is significantly higher than that of the Cap1 analog (3'-OMe-7mG) capped mRNA in different cell lines* (3T3-L1，Hela，JAWs， HEK293T and Huh7)，The 130 repetition experiment showed about a 1.5 times higher expression of LZCap AG(3'Acm) capped luciferase mRNA than the (3'-OMe-7mG) capped mRNA on average. Similar result is observed in mice. Protein expression level may vary slightly with different mRNA sequence.

Q4. Do you have more animal data?

The expression efficiency of LZCap AG(3'Acm) or LZCap AG(3'FMom) capped

GLuc encoding mRNA shows a higher in vivo expression relative to CapAG (3'-OMe-7mG) capped mRNAs in Cynomolgus Monkey and Pig. 

Q5. How about the innate immunogenicity of LZCap AG capped mRNAs ?

LZCapAG ( 3'Acm ) capped mRNAs show low innate immunogenicity. TLR8, TLR7, IL-1A and B play an important role in the immune response induced by uncapped RNA. The in vivo studies of LZCapped mRNA immunogenicity analysis showed that uncapped RNA caused significant changes in the transcription level of immune-related factors in mice. Both LZCap and 3'-OMe-7mG capped mRNAs display similar lower immune factor transcription level in mice after a single RNA-stimulated injection.

Q6. Did you do some safety test?

Yes, we did Cytotoxicity Test, Human Polymerase Inhibition Study and Ames Test.

1. There are no or little cytotoxicity observed for the nucleoside monomer of 3'-Acm-7mG（CC50＞10000nM）in 293T、Huh7、MRC5、THP1 and U87MG cells.
2. Human DNA polymerase ( α ,β , γ and Klenow) and mitochondrial RNA polymerase (hPOLRMT) activity inhibition studies show that 3'-Acm-7mG TP does not inhibit human DNA or RNA polymerase.
3. The bacterial reverse mutation test (Ames) detects associated genetic alterations, as well as genotoxic carcinogens in most rodents and humans. Ames test showed 3'-Acm-7mG has no genotoxicity.

Q7. Has this product been patented?

Yes. The patent has been granted in the USA.

Documents:

Safety Data Sheet

10684_MSDS_HB250708.pdf

Manuals

10684_Manual_Ver.EN20250613.pdf

Related Blog:

The Evolution and Importance of Cap Structures in mRNA Technology