Combinatorial library of chalcogen-containing lipidoids for intracellular delivery of genome-editing proteins

Li, Yamin; Yang, Tao; Yu, Yingjie; Shi, Nicola; Yang, Liu; Glass, Zachary; Bolinger, Justin; Finkel, Isaac James; Li, Wenhan; Xu, Qiaobing

doi:10.1016/j.biomaterials.2018.03.011

Cited by 67 publications

(102 citation statements)

References 54 publications

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“…Nitrilotriacetic acid group–containing amphiphilic lipidoids NTA‐EC16, NTA‐O16B, and NTA‐O17O (Figure B) were synthesized by reacting Nα,Nα ‐bis(carboxymethyl)‐ l ‐lysine head group with hydrophobic tails (EC16,[8c] O16B,[8b,d] and O17O[8a]; Figure S1, Supporting Information); the products were purified using a Teledyne ISCO Chromatography purification system. The structures of NTA‐lipidoids were confirmed by electrospray ionization mass spectrometry (ESI‐MS) and NMR analysis and the results are summarized in Table S1 and Figure S2 in the Supporting Information.…”

Section: Resultsmentioning

confidence: 99%

“…Our group has developed several combinatorial libraries of lipid‐like molecule (lipidoid)‐based nanoparticles (LNPs) for intracellular delivery applications; the effectiveness of the LNPs has been shown by us and others in both in vitro and in vivo studies. [8c,9] These cationic lipidoids contain amine head groups and the main driving force for cargo complexation is the electrostatic interaction between LNP carriers and anionic guest molecules (negatively charged proteins, mRNA, siRNA, pDNA, etc.). [8e,10] The possibility of using noncationic lipidoids and other types of supramolecular interactions for protein loading and delivery is explored in this study.…”

Section: Introductionmentioning

confidence: 99%

“…[8c,9] These cationic lipidoids contain amine head groups and the main driving force for cargo complexation is the electrostatic interaction between LNP carriers and anionic guest molecules (negatively charged proteins, mRNA, siRNA, pDNA, etc.). [8e,10] The possibility of using noncationic lipidoids and other types of supramolecular interactions for protein loading and delivery is explored in this study.…”

Section: Introductionmentioning

confidence: 99%

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Intracellular Delivery of His‐Tagged Genome‐Editing Proteins Enabled by Nitrilotriacetic Acid–Containing Lipidoid Nanoparticles

2018

Adv Healthcare Materials

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and metal oxide nanoparticles, [7] etc. have been reported for protein delivery purposes. Our group has developed several combinatorial libraries of lipid-like molecule (lipidoid)-based nanoparticles (LNPs) for intracellular delivery applications [8] ; the effectiveness of the LNPs has been shown by us and others in both in vitro and in vivo studies. [8c,9] These cationic lipidoids contain amine head groups and the main driving force for cargo complexation is the electrostatic interaction between LNP carriers and anionic guest molecules (negatively charged proteins, mRNA, siRNA, pDNA, etc.). [8e,10] The possibility of using noncationic lipidoids and other types of supramolecular interactions for protein loading and delivery is explored in this study.One of the most widely used protein purification processes takes advantage of the coordination interaction between the nickel-immobilized beads and the imidazole groups on the His-tagged proteins. Dowdy [11] and Zuber [12] developed nitrilotriacetic acid (NTA) moiety-containing peptide transduction domains and synthetic polymers complexed with divalent nickel ions for His-tagged protein/nanoparticle cargo binding and intracellular delivery. Inspired by their designs, we report here the synthesis of three types of NTA-containing lipidoids with different hydrophobic tail structures (NTA-EC16, NTA-O16B, and NTA-O17O; Figure 1) for protein delivery. As a proof of concept, nanoparticles were fabricated using active lipidoids containing NTA groups, together with divalent nickel ions and helper lipids (cholesterol, phospholipid, and macromolecular lipids; Figure 1B) for optimized intracellular delivery efficiencies. Histagged proteins including green fluorescent protein (GFP), Cre recombinase variant, and recently developed CRISPR (clustered regularly interspaced short palindromic repeat)-associated protein 9 (Cas9) nuclease were complexed with the nanoparticle formulations and their internalization efficiencies as well as genome-editing activities of Cre recombinase and Cas9 ribonucleoprotein (RNP) were studied ( Figure 1A). Results and Discussion Lipidoid Synthesis and Nanoparticle FabricationNitrilotriacetic acid group-containing amphiphilic lipidoids NTA-EC16, NTA-O16B, and NTA-O17O ( Figure 1B) were Protein-and peptide-based therapeutics with high tolerance and specificity along with low off-target effects and genetic risks have attracted tremendous attention over the last three decades. Herein, a new type of noncationic lipidoid nanoparticle (LNP) is reported for His-tagged protein delivery. Active lipidoids are synthesized by conjugating a nitrilotriacetic acid group with hydrophobic tails (EC16, O16B, and O17O) and nanoparticles are formulated in the presence of nickel ions and helper lipids (cholesterol, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, and 1,2-distearoyl-snglycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]). It is demonstrated that the newly developed LNPs are capable of delivering various His-tagged proteins including green fluore...

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Intracellular Delivery of His‐Tagged Genome‐Editing Proteins Enabled by Nitrilotriacetic Acid–Containing Lipidoid Nanoparticles

2018

Adv Healthcare Materials

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“…This screening has shed light on the structure-activity relationship between lipid structure and delivery efficacy. [34] Al ibrary of PEG-blockpoly(d,l-lactic-co-glycolic acid) (PEG-b-PLGA)-based cationic lipid-assisted nanoparticles (CLANs) that carry macrophagespecific promotor (CD68)-driving Cas9 expression plasmidw as recently reported. With the guidance of the sgRNA-targeting Ntn1 gene, the CLANs specifically edit the Ntn1 gene only in macrophages andr educe the expression of netrin1a nd improve typeIIdiabetes symptomsinv ivo (Figure3C).…”

Section: Physical Deliverymentioning

confidence: 97%

“…Other work has identified lipids from a library of synthetic cationic lipids that effectively deliver the Cas9 protein. This screening has shed light on the structure–activity relationship between lipid structure and delivery efficacy . A library of PEG‐block‐poly( d,l ‐lactic‐co‐glycolic acid) (PEG‐b‐PLGA)‐based cationic lipid‐assisted nanoparticles (CLANs) that carry macrophage‐specific promotor (CD68)‐driving Cas9 expression plasmid was recently reported.…”

Section: Delivery Of Crispr/cas9mentioning

confidence: 99%

Delivery of CRISPR/Cas9 by Novel Strategies for Gene Therapy

et al. 2018

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Precise editing of the genome of a living body is a goal pursued by scientists in many fields. In recent years, CRISPR (clustered regularly interspaced short palindromic repeat)/Cas (CRISPR‐associated) genome‐editing systems have become a revolutionary toolbox for gene editing across various species. However, the low transfection efficiency of the CRISPR/Cas9 system to mammalian cells in vitro and in vivo is a big obstacle hindering wide and deep application. In this review, recently developed delivery strategies for various CRISPR/Cas9 formulations and their applications in treating gene‐related diseases are briefly summarized. This review should inspire others to explore more efficient strategies for CRISPR system delivery and gene therapy.

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Simple and Efficient Targeted Intracellular Protein Delivery with Self‐Assembled Nanovehicles for Effective Cancer Therapy

Long

Zeng

et al. 2019

Adv Funct Materials

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Protein therapy offers promising prospects for the treatment of various important diseases, thus it is highly desirable to develop a robust carrier that can deliver active proteins into cells. The development of a novel protein delivery platform based on the self‐assembly of multiarmed amphiphilic cyclodextrins (CDEH) is reported. CDEH can self‐assemble into nanoparticles in aqueous solution and achieve superior encapsulation of protein (loading capacity > 30% w/w) simply by mixing with protein solution without introducing any subsequent cumbersome steps that may inactivate proteins. More importantly, CDEH nanovehicles can be easily further modified with various targeting groups based on host–guest complexation. Using saporin as a therapeutic protein, AS1411‐aptamer‐modified CDEH nanovehicles can preferentially accumulate in tumors and efficiently inhibit tumor growth in a MDA‐MB‐231 xenograft mouse model. Moreover, folate‐targeted CDEH nanovehicles can also deliver Cas9 protein and Plk1‐targeting sgRNA into Hela cells, leading to 47.1% gene deletion and 64.1% Plk1 protein reduction in HeLa tumor tissue, thereby effectively suppressing the tumor progression. All these results indicate the potential of targeted CDEH nanovehicles in intracellular protein delivery for improving protein therapeutics.

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Combinatorial library of chalcogen-containing lipidoids for intracellular delivery of genome-editing proteins

Cited by 67 publications

References 54 publications

Intracellular Delivery of His‐Tagged Genome‐Editing Proteins Enabled by Nitrilotriacetic Acid–Containing Lipidoid Nanoparticles

Intracellular Delivery of His‐Tagged Genome‐Editing Proteins Enabled by Nitrilotriacetic Acid–Containing Lipidoid Nanoparticles

Delivery of CRISPR/Cas9 by Novel Strategies for Gene Therapy

Simple and Efficient Targeted Intracellular Protein Delivery with Self‐Assembled Nanovehicles for Effective Cancer Therapy

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