Active Delivery of CRISPR System Using Targetable or Controllable Nanocarriers

Abstract

Among programmable nuclease‐based genome editing tools, the clustered regularly interspaced short palindromic repeats (CRISPR) system with accuracy and the convenient operation is most promising to be applied in gene therapy. The development of effective delivery carriers for the CRISPR system is the major premise to achieve practical applications. Although many nanocarrier‐mediated deliveries have been reported to be safer and cheaper over the physical and viral delivery, the accumulation at disease sites or controllability with the spatial or temporal resolution are still desired on nanocarriers to reduce side effects and off‐target from the CRISPR system. Therefore, the targetable and controllable nanocarriers to actively deliver the CRISPR system are summarized. The cell or even organ selective nanocarriers are introduced first, followed by the discussion of nanocarriers controlled by biochemical or physical signals. At last, the potential challenges faced by existing nanocarriers are discussed.

Nanomaterial-assisted CRISPR gene-engineering – A hallmark for triple-negative breast cancer therapeutics advancement

“…It deals with various techniques for diagnosing, imaging, controlling, and delivering several types of payloads to the desired targeted site [ 39 ]. It supports the distribution of CRISPR/Cas at high efficiency and lesser toxicity that would overwhelm physiological obstacles [ 47 ]. The encapsulation via nano-based cargo precludes the susceptible CRISPR complex from deterioration intervened by proteases and nucleases in biological fluids matrix [ 51 ].…”

“…However, safe and direct CRISPR/Cas cargo delivery at the targeted site [ 41 ], can be achieved via physical methods such as electroporation, microinjection, hydrodynamic, and physical stimulus method [ 41 , 45 ], and virus-mediated delivery [ 46 ] method. Both methods of the CRISPR payload delivery are highly effective in the wet-laboratory, nonetheless they can cause impairment due to the destruction of cell membranes and seditious reaction encouraged by immunogenicity, higher off-target impact, limited capacity of packaging, and high mass production cost [ 47 , 48 ]. Additionally, it may be shoddier that the ectopic chromosomal incorporation facilitated thru virus that can disturb the regulatory sequence of oncogenes or tumour suppressor genes leading to either secondary melanoma or more violent metastasis.…”

Exploring nano-enabled CRISPR-Cas-powered strategies for efficient diagnostics and treatment of infectious diseases

“…Natural extracellular vesicles, as well as synthetic nanoparticles based on liposomes, polymers, or polypeptides, have been used to deliver CRISPR/Cas9 [175]. Targetable unit manipulation on nanocarriers to accomplish specific gene delivery is a promising technique that is being extensively used in gene therapy to reduce risk factors on body cells [176]. Encapsulated nanocarrier protects susceptible CRISPR materials from degradation in biological fluids by nucleases and proteases.…”

Effective Genome Editing Using CRISPR‐Cas9 Nanoflowers

“…Cellular uptake is a prerequisite process for Cas9 RNP to perform gene editing in nucleus [3,39] , Therefore, the intracellular delivery behaviors of Cas9-NF were next investigated. Different FITC-labelled Cas9 formulations including free Cas9, Cas9-NF, Cas9-NP (Cas9-NP-NPC hereinafter) were incubated with A549 cells, a human pulmonary carcinoma cell line used for the following in vitro and in vivo tumor studies.…”

“…CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats) systems have emerged as a powerful genome-editing tool to treat genetic diseases, directed by a single guide RNA (sgRNA) to cause DNA strand breaks. [1][2][3] In order to deliver the Cas9 and sgRNA ribonucleoprotein (RNP) complex to the nucleus for gene editing, many delivery systems have been developed including viral constructs, [4,5] mesoporous silica nanoparticles, [6] metal organic frameworks, [7,8] gold nanoparticles, [9,10] liposomes, [11,12] extracellular vesicles, [13,14] DNA-based materials, [15] and polymers. [16,17] These delivery systems have shown great promise but also left room for improvements due to immunogenicity, poor stability and toxicity.…”