The manipulation of cultured mammalian cells by the delivery of exogenous macromolecules is one of the cornerstones of experimental cell biology. Although the transfection of cells with DNA expressions constructs that encode proteins is routine and simple to perform, the direct delivery of proteins into cells has many advantages. For example, proteins can be chemically modified, assembled into defined complexes and subject to biophysical analyses prior to their delivery into cells. Here, we review new approaches to the injection and electroporation of proteins into cultured cells. In particular, we focus on how recent developments in nanoscale injection probes and localized electroporation devices enable proteins to be delivered whilst minimizing cellular damage. Moreover, we discuss how nanopore sensing may ultimately enable the quantification of protein delivery at single-molecule resolution.
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Cover Image
Cover Image
The cover shows a metaphorical representation of the anti-CRISPR AcrIIA6, represented as handcuffs, sequestering two Streptococcus thermophilus CRISPR1-Cas9 (St1Cas9) molecules at a time and preventing conformational changes associated with DNA recognition and binding. In the absence of AcrIIA6, St1Cas9 tightly binds to its target DNA, and can proceed to target cleavage. For further information, see the article by Hardouin and Goulet in this issue (pp. 507–516). This cover artwork has been made by Beata Edyta Mierzwa (www.BeataScienceArt.com).
Methods for protein delivery into cells: from current approaches to future perspectives
Chalmers Chau, Paolo Actis, Eric Hewitt; Methods for protein delivery into cells: from current approaches to future perspectives. Biochem Soc Trans 29 April 2020; 48 (2): 357–365. doi: https://doi.org/10.1042/BST20190039
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