RNA-Guided Human Genome Engineering via Cas9

Mali, Prashant; Yang, Luhan; Esvelt, Kevin M.; Aach, John; Güell, Marc; DiCarlo, James E.; Norville, Julie E.; Church, George M.

doi:10.1126/science.1232033

Cited by 8,033 publications

(6,921 citation statements)

References 48 publications

Supporting

Mentioning

6,796

Contrasting

Unclassified

Order By: Relevance

“…In the CRISPR‐Cas9 system, the Cas9, an endonuclease, is directed by a single guide RNA (sgRNA) to cause double‐strand break of target DNA sequences with high specificity,2 allowing for much greater ease of construction of knockout reagents than other methods for genetic modification 3, 4, 5. However, it is very challenging for the delivery of the CRISPR‐Cas9 system into cells or tissues because the plasmid encoding both Cas9 and sgRNA has strong negative charges and large size (usually exceeds 10 000 bp).…”

Section: Introductionmentioning

confidence: 99%

Genome Editing for Cancer Therapy: Delivery of Cas9 Protein/sgRNA Plasmid via a Gold Nanocluster/Lipid Core–Shell Nanocarrier

et al. 2017

View full text Add to dashboard Cite

The type II bacterial clustered, regularly interspaced, short palindromic repeats (CRISPR)‐Cas9 (CRISPR‐associated protein) system (CRISPR‐Cas9) is a powerful toolbox for gene‐editing, however, the nonviral delivery of CRISPR‐Cas9 to cells or tissues remains a key challenge. This paper reports a strategy to deliver Cas9 protein and single guide RNA (sgRNA) plasmid by a nanocarrier with a core of gold nanoclusters (GNs) and a shell of lipids. By modifying the GNs with HIV‐1‐transactivator of transcription peptide, the cargo (Cas9/sgRNA) can be delivered into cell nuclei. This strategy is utilized to treat melanoma by designing sgRNA targeting Polo‐like kinase‐1 (Plk1) of the tumor. The nanoparticle (polyethylene glycol‐lipid/GNs/Cas9 protein/sgPlk1 plasmid, LGCP) leads to >70% down‐regulation of Plk1 protein expression of A375 cells in vitro. Moreover, the LGCP suppresses melanoma progress by 75% on mice. Thus, this strategy can deliver protein‐nucleic acid hybrid agents for gene therapy.

show abstract

Section: Introductionmentioning

confidence: 99%

Genome Editing for Cancer Therapy: Delivery of Cas9 Protein/sgRNA Plasmid via a Gold Nanocluster/Lipid Core–Shell Nanocarrier

et al. 2017

View full text Add to dashboard Cite

show abstract

“…CRISPR/Cas9 has attracted increasing attention since the report of its successful use in mammalian cells 36, 37 or human cells 37, 38. This approach has made gene editing easier and more efficient than the previous gene editing tools, such as zinc finger nuclease (ZFN) and transcription activator‐like effector nucleases (TALEN) 39.…”

Section: Discussionmentioning

confidence: 99%

Expression of PTEN‐long mediated by CRISPR/Cas9 can repress U87 cell proliferation

Fang

Wang

et al. 2017

J Cellular Molecular Medi

View full text Add to dashboard Cite

PTEN is a tumour suppressor that is frequently mutated in a variety of cancers. Hence, PTEN has significant potential as a therapeutic molecule. PTEN‐long is an alternative translation variant, with an additional 173 amino acids added to the N‐terminal of the canonical PTEN when CUG of the mRNA is utilized as the start codon. PTEN‐long is secreted into serum and can re‐enter cells throughout the body. One of the major barriers for gene therapy is to efficiently and specifically deliver DNA or RNA material to target cells. As an alternative approach, if a therapeutic protein can be directly delivered to target cell of interest, it should theoretically function well within the cells, particularly for genes that are deficiently expressed in vivo. Most therapeutic proteins are incapable of efficiently permeating the cell membrane. In this study, we have employed CRISPR/Cas9 gene editing tool combined with single‐stranded template to edit CTG of PTEN‐long to ATG in the genome. Two guide RNAs close to CTG site were found to have similar efficiency in driving PTEN‐long expression. Furthermore, we detected PTEN‐long expression in transfected whole‐cell lysate and in concentrated culture media in Western blot. Interestingly, the culture media of PTEN‐long expression can reduce Akt phosphorylation level and repress U87 cell proliferation compared to wild‐type U87 or control media. Taken together, PTEN‐long driven by CRISPR/Cas9 imports and exports cells and represses nearby cell proliferation, indicating the PTEN‐long generated by CRISPR/Cas9 has potential to be an alternative strategy for PTEN gene therapy.

show abstract

“…CRISPR is used in mouse and human cells, fuelling rapid uptake of the technique by researchers [11][12][13] .…”

Section: January 2013mentioning

confidence: 99%

CRISPR, the disruptor

Ledford¹

2015

Nature

309

188

View full text Add to dashboard Cite

RNA-Guided Human Genome Engineering via Cas9

Cited by 8,033 publications

References 48 publications

Genome Editing for Cancer Therapy: Delivery of Cas9 Protein/sgRNA Plasmid via a Gold Nanocluster/Lipid Core–Shell Nanocarrier

Genome Editing for Cancer Therapy: Delivery of Cas9 Protein/sgRNA Plasmid via a Gold Nanocluster/Lipid Core–Shell Nanocarrier

Expression of PTEN‐long mediated by CRISPR/Cas9 can repress U87 cell proliferation

CRISPR, the disruptor

Contact Info

Product

Resources

About