Cas9 RNP可以通过在 Lonza 4D Nucleofector系统 或 ThermoFisher Neon转染系统 中的电穿孔引入免疫细胞，以实现高基因敲除效率和细胞活力。 同时，多个不同特异性的Cas9 RNP可以在单个反应中传递，以同时靶向多个基因组位点。

Learn the basics of CRISPR ribonucleoprotein (RNP) preparation and delivery for your next gene editing experiment.

This user manual describes how to use GenScript’s CRISPR Ribonucleoprotein (RNP) products for targeted genome editing. The RNP system is comprised of a Cas9 protein, which creates double strand breaks in the target genome, and two synthetic oligos: CRISPR-targeting RNA (crRNA) and trans-activating crRNA (tracrRNA).

This user manual describes how to use GenScript’s CRISPR Ribonucleoprotein (RNP) products for targeted genome editing. The RNP system is comprised of a Cas9 protein, which creates double strand breaks in the target genome, and two synthetic oligos: CRISPR-targeting RNA (crRNA) and trans-activating crRNA (tracrRNA).

2024年11月27日 · In this article, we provide detailed protocols to perform efficient gene knock-out (KO) or genomic deletion in a small number of HKs without clonal selection of edited cells. By nucleofecting ribonucleoprotein complexes, we efficiently generate KO cells as well as deletion of specific genomic regions.

2025年2月25日 · Several commercial sources of Cas9 protein suitable for zygote electroporation with ribonucleoprotein (RNP) also exist. In Basic Protocol 2 we describe the steps for designing the oligonucleotide repair template needed to effect precise genome sequence changes.

Genome editing by the delivery of pre-assembled Cas9 ribonucleoproteins (Cas9 RNP) is an increasingly popular approach for cell types that are difficult to manipulate genetically by the conventional plasmid and viral methods. Cas9 RNP editing is robust, precise, capable of multiplexing, and free of genetic materials.

Direct delivery of CRISPR/Cas9 system as a ribonucleoprotein (RNP) complex consisting of Cas9 protein and single guide RNA (sgRNA) has emerged as a powerful and widespread method for genome editing due to its advantages of transient genome editing and reduced off-target effects.

co-CRISPR RNP Injection Mix Protocol Stefani Giacopazzi (adapted from Dernburg protocol and Zahler protocol), Bhalla Lab, Nov. 2017 Resuspend target-gRNA and dpy-10-gRNA in IDT duplex buffer Add 10ul duplex buffer to each to resuspend to 200uM gRNA **Keep resuspended RNAs on ice to prevent degradagtion.

Table 1: RNP preparation for 1 Monkey (570ul) - 2x concentrated, as it will be diluted 1:1 in the Delivery Mix prep) Table 2: Delivery Mix preparation for Monkey #6