Efficient intracellular delivery of native proteins

Diego S D'Astolfo; Romina J Pagliero; Anita Pras; Wouter R Karthaus; Hans Clevers; Vikram Prasad; Robert Jan Lebbink; Holger Rehmann; Niels Geijsen

doi:10.1016/j.cell.2015.03.028

Efficient intracellular delivery of native proteins

Diego S D'Astolfo, Romina J Pagliero, Anita Pras, Wouter R Karthaus, Hans Clevers, Vikram Prasad, Robert Jan Lebbink, Holger Rehmann, Niels Geijsen

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

261 Citations (Scopus)

Abstract

Modulation of protein function is used to intervene in cellular processes but is often done indirectly by means of introducing DNA or mRNA encoding the effector protein. Thus far, direct intracellular delivery of proteins has remained challenging. We developed a method termed iTOP, for induced transduction by osmocytosis and propanebetaine, in which a combination of NaCl hypertonicity-induced macropinocytosis and a transduction compound (propanebetaine) induces the highly efficient transduction of proteins into a wide variety of primary cells. We demonstrate that iTOP is a useful tool in systems in which transient cell manipulation drives permanent cellular changes. As an example, we demonstrate that iTOP can mediate the delivery of recombinant Cas9 protein and short guide RNA, driving efficient gene targeting in a non-integrative manner.

Original language	English
Pages (from-to)	674-90
Number of pages	17
Journal	Cell
Volume	161
Issue number	3
DOIs	https://doi.org/10.1016/j.cell.2015.03.028
Publication status	Published - 23 Apr 2015

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title = "Efficient intracellular delivery of native proteins",

abstract = "Modulation of protein function is used to intervene in cellular processes but is often done indirectly by means of introducing DNA or mRNA encoding the effector protein. Thus far, direct intracellular delivery of proteins has remained challenging. We developed a method termed iTOP, for induced transduction by osmocytosis and propanebetaine, in which a combination of NaCl hypertonicity-induced macropinocytosis and a transduction compound (propanebetaine) induces the highly efficient transduction of proteins into a wide variety of primary cells. We demonstrate that iTOP is a useful tool in systems in which transient cell manipulation drives permanent cellular changes. As an example, we demonstrate that iTOP can mediate the delivery of recombinant Cas9 protein and short guide RNA, driving efficient gene targeting in a non-integrative manner.",

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T1 - Efficient intracellular delivery of native proteins

AU - D'Astolfo, Diego S

AU - Pagliero, Romina J

AU - Pras, Anita

AU - Karthaus, Wouter R

AU - Clevers, Hans

AU - Prasad, Vikram

AU - Lebbink, Robert Jan

AU - Rehmann, Holger

AU - Geijsen, Niels

PY - 2015/4/23

Y1 - 2015/4/23

N2 - Modulation of protein function is used to intervene in cellular processes but is often done indirectly by means of introducing DNA or mRNA encoding the effector protein. Thus far, direct intracellular delivery of proteins has remained challenging. We developed a method termed iTOP, for induced transduction by osmocytosis and propanebetaine, in which a combination of NaCl hypertonicity-induced macropinocytosis and a transduction compound (propanebetaine) induces the highly efficient transduction of proteins into a wide variety of primary cells. We demonstrate that iTOP is a useful tool in systems in which transient cell manipulation drives permanent cellular changes. As an example, we demonstrate that iTOP can mediate the delivery of recombinant Cas9 protein and short guide RNA, driving efficient gene targeting in a non-integrative manner.

AB - Modulation of protein function is used to intervene in cellular processes but is often done indirectly by means of introducing DNA or mRNA encoding the effector protein. Thus far, direct intracellular delivery of proteins has remained challenging. We developed a method termed iTOP, for induced transduction by osmocytosis and propanebetaine, in which a combination of NaCl hypertonicity-induced macropinocytosis and a transduction compound (propanebetaine) induces the highly efficient transduction of proteins into a wide variety of primary cells. We demonstrate that iTOP is a useful tool in systems in which transient cell manipulation drives permanent cellular changes. As an example, we demonstrate that iTOP can mediate the delivery of recombinant Cas9 protein and short guide RNA, driving efficient gene targeting in a non-integrative manner.

U2 - 10.1016/j.cell.2015.03.028

DO - 10.1016/j.cell.2015.03.028

M3 - Article

C2 - 25910214

SN - 0092-8674

VL - 161

SP - 674

EP - 690

JO - Cell

JF - Cell

IS - 3

ER -

Efficient intracellular delivery of native proteins

Abstract

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