Antibody-targeted chromatin enables effective intracellular delivery and functionality of CRISPR/Cas9 expression plasmids

Nucleic Acids Research
Tobias KillianUlrich Brinkmann

Abstract

We report a novel system for efficient and specific targeted delivery of large nucleic acids to and into cells. Plasmid DNA and core histones were assembled to chromatin by salt gradient dialysis and subsequently connected to bispecific antibody derivatives (bsAbs) via a nucleic acid binding peptide bridge. The resulting reconstituted vehicles termed 'plasmid-chromatin' deliver packaged nucleic acids to and into cells expressing antigens that are recognized by the bsAb, enabling intracellular functionality without detectable cytotoxicity. High efficiency of intracellular nucleic acid delivery is revealed by intracellular expression of plasmid encoded genes in most (∼90%) target cells to which the vehicles were applied under normal growth/medium conditions in nanomolar concentrations. Specific targeting, uptake and transgene expression depends on antibody-mediated cell surface binding: plasmid chromatin of identical composition but with non-targeting bsAbs or without bsAbs is ineffective. Examples that demonstrate applicability, specificity and efficacy of antibody-targeted plasmid chromatin include reporter gene constructs as well as plasmids that enable CRISPR/Cas9 mediated genome editing of target cells.

References

Jan 1, 1989·Methods in Enzymology·D Rhodes, R A Laskey
May 14, 1993·Science·R C Mulligan
Jul 1, 1997·Nature Biotechnology·L Huang, S Li
Oct 4, 2000·Proceedings of the National Academy of Sciences of the United States of America·D BalickiH N Lode
Dec 6, 2001·Current Drug Targets·M Johnson-Saliba, D A Jans
May 29, 2002·Proceedings of the National Academy of Sciences of the United States of America·Danuta BalickiErnest Beutler
May 2, 2003·Journal of Biomedicine & Biotechnology·Andreas G. Schätzlein
May 3, 2003·Nature Reviews. Genetics·Clare E ThomasMark A Kay
Jul 29, 2003·Journal of Cancer Research and Clinical Oncology·Fei-Han DaiJian-Ren Gu
Sep 19, 2003·Biochemical and Biophysical Research Communications·Annick LucasGwendal Lazennec
Oct 25, 2003·Journal of Cell Science·Elana Hariton-GazalAbraham Loyter
Oct 29, 2003·Journal of Biotechnology·Iratxe PueblaWalter Low
Nov 25, 2003·The Journal of Immunology : Official Journal of the American Association of Immunologists·Barbara SpiesHermann Wagner
Feb 12, 2004·Methods in Enzymology·Pamela N DyerKarolin Luger
Mar 6, 2004·Science·Volker BrinkmannArturo Zychlinsky
Jul 23, 2004·Current Biology : CB·Craig L Peterson, Marc-André Laniel
Dec 2, 2004·Journal of Molecular Biology·Joseph RosenbluhAbraham Loyter
Jun 7, 2005·Expert Opinion on Biological Therapy·Shin-ichi MuramatsuKeiya Ozawa
Mar 1, 2007·Molecular Therapy : the Journal of the American Society of Gene Therapy·Kylie M WagstaffDavid A Jans
Jun 19, 2007·Biotechnology Progress·Hee Jung JungHyung Joon Cha
Jul 4, 2007·Nature Reviews. Genetics·Reinhard WaehlerDavid T Curiel
Oct 24, 2007·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jon A Wolff, David B Rozema
Mar 22, 2008·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Kylie M WagstaffDavid A Jans
Jan 17, 2009·Advanced Drug Delivery Reviews·Niek SandersJoseph Demeester
Sep 8, 2009·Journal of the Royal Society, Interface·Christopher L Grigsby, Kam W Leong
Nov 13, 2009·Gene Therapy·S Nayak, R W Herzog
Sep 8, 2010·International Journal of Pharmaceutics·Anne SchneeweissSebastian Ulbert
Nov 17, 2010·Journal of Controlled Release : Official Journal of the Controlled Release Society·Amir K VarkouhiHidde J Haisma
Apr 7, 2011·Nature Reviews. Genetics·Mark A Kay
Apr 19, 2011·Nature Reviews. Genetics·Federico Mingozzi, Katherine A High
Apr 20, 2011·International Journal of Biometeorology·Dusan FialaGerd Jendritzky
May 4, 2011·Proceedings of the National Academy of Sciences of the United States of America·Silke MetzUlrich Brinkmann
Dec 14, 2011·The Biochemical Journal·Alexander K HaasUlrich Brinkmann
Jan 27, 2012·Current Opinion in Structural Biology·Jeffrey A Speir, John E Johnson
Jul 11, 2012·Journal of Drug Delivery·José M MorachisAdah Almutairi
Sep 13, 2012·Molecular Therapy : the Journal of the American Society of Gene Therapy·Robert C MünchChristian J Buchholz

❮ Previous
Next ❯

Citations

Nov 5, 2019·Cancer Gene Therapy·Shuai Zhen, Xu Li
Jul 4, 2019·Current Gene Therapy·Cia-Hin Lau, Chung Tin
Apr 22, 2020·Methods : a Companion to Methods in Enzymology·Mohammad Mijanur Rahman, Trygve O Tollefsbol
Nov 24, 2019·Advanced Drug Delivery Reviews·Cong-Fei XuJun Wang
Feb 2, 2022·Biomaterials Science·Yi LinUlrich Lächelt

❮ Previous
Next ❯

Methods Mentioned

BETA
transfection
chemical modification
salt gradient dialysis
size exclusion chromatography
flow cytometry
fluorescence
electrophoresis
microscale thermophoresis
peptide display
confocal microscopy

Software Mentioned

MST
2bind

Related Concepts

Related Feeds

Antibody Engineering

Antibody engineering technologies are constantly advancing to improve the clinical effectiveness of monoclonal and bispecific antibodies. Discover the latest research on Antibody Engineering here.

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.