GenLNP-A01-eSpCas9 mRNA(m1Ψ)-TRAC sgRNA

The LNP formulation loaded with eSpCas9 mRNA (m1Ψ) and TRAC sgRNA (at the molar ratio of 1:10) is prepared using generic LNP formulation with main ionizable lipid of ALC0315. The payload eSpCas9 mRNA (m1Ψ) has all the U bases 100% modified with N1-methyl-pseudoUridine. HPLC purified SafeEdit sgRNA with modifications targeting the first exon of the constant chain of the TCRα gene (TRAC), which enhances CAR-T cell potency and persistence by utilizing the endogenous transcriptional control of TCR gene. This formulation is a good control for testing if ALC0315 LNP is a good potential formulation for your CRISPR gene know out product under research in your in vitro and in vivo experiment model.

价格	￥2000 在线订购包邮(港澳台除外)
产品编号	RP-A00019-50
规格	0.05 mg 大单询价
数量
参考货期


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产品属性质控参数应用指导图例

Description

The LNP formulation loaded with eSpCas9 mRNA (m1Ψ) and TRAC sgRNA is prepared using generic LNP formulation with main ionizable lipid of ALC0315.

The payload eSpCas9 mRNA (m1Ψ) has all the U bases 100% modified with N1-methyl-pseudouridine (m1Ψ). HPLC purified SafeEdit sgRNA with modifications targeting the first exon of the constant chain of the TCRα gene (TRAC), which enhances CAR-T cell potency and persistence by utilizing the endogenous transcriptional control of TCR gene.

The eSpCas9 mRNA and TRAC sgRNA are loaded at a molar ratio of 1:10 in this LNP formulation.

This formulation is a good control for testing if ALC0315 LNP formulation is a good potential formulation for your CRISPR gene know out product under research in your in vitro and in vivo experiment model.

产品属性

Form	Liquid
Concentration	0.15mg/mL
Full mRNA length	4471nt
Full mRNA Molecular Weight	1457890
Storage buffer	Tris-HCl/Sucrose
Storage condition	Store at -80°C for long term. Avoid freeze thaw, only thaw once before use.

质控参数

Appearance	Clear and free of foreign particles
Encapsulation Efficiency	> 85%
Endotoxin	< 4 EU/ml
Particle Size	50-100 nm
pH	7.4 ± 0.5
Polydispersity Index	< 0.2
Zeta Potential	± 15.0 mV

应用指导

Guide for LNP-CRISPR gene knockout (KO) operation of Jurkat cells:

1. Cell culture
- Change the culture medium every 2-3 days.
- Passage the cells 2-3 times per week. The recommended cell passage ratio is 1:5 - 1:6.
- Maintain the cell density during culture at 2×105 - 1.5×106 cells/ml.

2. LNP transfection (24-well plate)
- On the day of transfection, perform cell counting to determine the culture density. Add 0.5 ml of complete growth medium to each well, and plate with 1 x 105 cells. On the day of transfection, the cell density should reach ~80% confluence.
- Directly add LNP containing 0.5μg Cas9 mRNA & sgRNA to each cell well, and gently shake the culture plate back and forth to mix.
- After transfection, incubate the cells in a 37°C CO2 cell culture incubator for 2-3 days for gene knockout analysis.

图例

For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.