CRISPR-Cas9 technology has revolutionized the field of genome editing, offering unprecedented precision and efficiency in targeted genetic modifications. One of the critical components of CRISPR-mediated genome editing is the delivery of single guide RNA (sgRNA) to the target cells. Optimizing sgRNA delivery methods is crucial for maximizing the efficiency and specificity of CRISPR editing. In this blog post, we will explore various strategies for enhancing sgRNA delivery, thereby improving CRISPR efficiency.

Cell transfection techniques are commonly used in research to effectively introduce exogenous DNA, RNA, or proteins into cells for purposes such as gene editing and protein expression. Electroporation and lipofection are two common transfection methods that play important roles in the field of cell transfection. Electroporation utilizes brief high-voltage pulses to increase the permeability of cell membranes, allowing exogenous DNA or RNA to enter the cells, while lipofection involves the formation of complexes between lipids and exogenous DNA or RNA, which then bind to the cell membrane and are engulfed by the cell, facilitating the entry of exogenous molecules into the cytoplasm. This study aims to compare the knockout efficiency of these two transfection methods for Cas9 protein and mRNA in Hek 293T and HeLa cells to reveal the impact of different transfection methods on gene editing efficiency in two cell types.

In October 2019, Andrew Anzalone, a postdoctoral fellow in David Liu's lab, and colleagues unveiled a groundbreaking innovation in genetic engineering known as Prime Editing, in a publication featured in Nature. This development represents a significant stride forward in the realm of genetic manipulation, offering a highly precise and versatile method that addresses several limitations encountered with conventional CRISPR-Cas9 systems. Let's delve into the essence of Prime Editing, exploring its functionality andseveral advantages.

A few weeks back we presented to you data on the mutation efficiency of sgRNAs produced with our New Modality Enzymatic Convergent Assembly (nMECA) technology. There was one bit we did not cover back then. We would like to bring back those data into a different light. Back then, we also reported the use of fluorescent sgRNA. nMECA not only allows us to produce extra-long sgRNAs and pegRNAs with an unprecedentedly high purity in the market (>90%), but also to deliver to you these products attached to any fluorescent label of your choice.

This week's post we cover what our technology new Modality Enzymatic Convergent Assembly (nMECA) is and how we accomplished long sequences (>180 nt) with purities of >90%. Even more importantly, why this is an advantage for your research.

Despite the big, immediate success that CRISPR-Cas gene editing initially had, providing unprecedented efficiency and precision, there was still a lot to be done. Soon researchers realized that this new system was not flawless: off-site mutations occurred a bit too often, a problem that immediately hindered its way towards clinical applications. The cures of genetic diseases from the root still had wait.