With the rise of new powerful genome engineering technologies, such as CRISPR/Cas9, cell models can be engineered effectively to accelerate basic and disease research. The most critical step in this procedure is the efficient delivery of foreign nucleic acids into cells by cellular transfection. Since the vectors encoding the components necessary for CRISPR/Cas genome engineering are always large (9–19 kb), they result in low transfection efficiency and cell viability, and thus subsequent selection or purification of positive cells is required. To overcome those obstacles, we here show a non-toxic and non-viral delivery method that increases transfection efficiency (up to 40-fold) and cell viability (up to 6-fold) in a number of hard-to-transfect human cancer cell lines and primary blood cells. At its core, the technique is based on adding exogenous small plasmids of a defined size to the transfection mixture.

随着新的强大基因组工程技术（例如CRISPR / Cas9）的兴起，可以有效地对细胞模型进行工程设计，以加速基础研究和疾病研究。该过程中最关键的步骤是通过细胞转染将外来核酸有效地递送到细胞中。由于编码CRISPR / Cas基因组工程必需成分的载体总是很大（9–19 kb），因此它们导致低转染效率和细胞活力，因此需要随后选择或纯化阳性细胞。为了克服这些障碍，我们在这里展示了一种无毒，无病毒的递送方法，可以在许多难以转染的人类癌症中提高转染效率（最高40倍）和细胞活力（最高6倍）。细胞系和原代血细胞。本质上，