Knockout Rats via Embryo Microinjection of Zinc-Finger Nucleases
It is demonstrated that a single injection of DNA or messenger RNA encoding ZFNs into the one-cell rat embryo leads to a high frequency of animals carrying 25 to 100% disruption at the target locus.
New fusion protein systems designed to give soluble expression in Escherichia coli.
- G. Davis, C. Elisee, D. Newham, R. Harrison
- BiologyBiotechnology and Bioengineering
- 20 November 1999
Three native E. coli proteins-NusA, GrpE, and bacterioferritin (BFR)-were studied in fusion proteins expressed in E. coli for their ability to confer solubility on a target insoluble protein at the…
High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases
- Fuqiang Chen, Shondra M. Pruett-Miller, G. Davis
- BiologyNature Methods
- 1 September 2011
Methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: targeted point mutation, targeted genomic deletion of up to 100 kb and targeted insertion of small genetic elements concomitant with large genomic deletions.
Functional genomics, proteomics, and regulatory DNA analysis in isogenic settings using zinc finger nuclease-driven transgenesis into a safe harbor locus in the human genome.
- Russell Dekelver, Vivian M. Choi, F. Urnov
- BiologyGenome Research
- 1 May 2010
The use of designed zinc finger nucleases (ZFNs) for efficient transgenesis without drug selection into the PPP1R12C gene, a "safe harbor" locus known as AAVS1, allows bona fide isogenic settings for high-throughput functional genomics, proteomics, and regulatory DNA analysis in essentially any transformed human cell type and in primary cells.
Actin and dynamin2 dynamics and interplay during clathrin-mediated endocytosis
- A. Grassart, Aaron T. Cheng, D. Drubin
- BiologyJournal of Cell Biology
- 9 June 2014
Actin assembly influences the precise temporal and quantitative recruitment of dynamin2 to sites of clathrin-mediated endocytosis.
Targeted activation of diverse CRISPR-Cas systems for mammalian genome editing via proximal CRISPR targeting
- Fuqiang Chen, Xiao Ding, Yongmei Feng, Timothy Seebeck, Yanfang Jiang, G. Davis
- BiologyNature Communications
- 7 April 2017
By manipulating local chromatin structures, inhibited Cas9s can have their activity restored in human cells, providing a novel strategy to enable use of diverse otherwise inactive CRISPR–Cas systems for genome-editing applications and a potential path to modulate the impact of chromatin microenvironments on genome modification.
Recombinant production and purification of novel antisense antimicrobial peptide in Escherichia coli.
- C. Haught, G. Davis, R. Subramanian, K. Jackson, R. Harrison
- BiologyBiotechnology and Bioengineering
- 5 January 1998
The yield of the fusion protein after expression by the cells was high, and the percentage recovery of the P2 peptide in the inclusion bodies was relatively low, which appears to be due to losses in the cyanogen bromide digestion step.
High-efficiency genome editing via 2A-coupled co-expression of fluorescent proteins and zinc finger nucleases or CRISPR/Cas9 nickase pairs
- K. Duda, Lindsey A. Lonowski, M. Frödin
- Biology, ChemistryNucleic Acids Research
- 21 April 2014
A broadly applicable and versatile method for increasing editing efficiencies by targeted endonucleases and clustered regularly interspaced short palindromic repeats, including CRISPR/Cas9 mutant nickase pairs, which exhibit low off-target mutagenesis compared to wild-type Cas9.
Enhancing the genome editing toolbox: genome wide CRISPR arrayed libraries
- E. Metzakopian, Alexander J. Strong, A. Bradley
- BiologyScientific Reports
- 22 May 2017
This is the first individually cloned CRISPR-Cas9 genome wide arrayed sgRNA libraries covering 17,166 human and 20,430 mouse genes at a complexity of 34,332 sgRNAs for human and 40,860 sg RNAs for the mouse genome, providing an organized comprehensive gene editing toolbox of considerable scientific value.
Improving CRISPR-Cas9 Genome Editing Efficiency by Fusion with Chromatin-Modulating Peptides.
- Xiao Ding, Timothy Seebeck, Yongmei Feng, Yanfang Jiang, G. Davis, Fuqiang Chen
- BiologyThe CRISPR Journal
- 1 February 2019
Modification of the widely used Streptococcus pyogenes Cas9 by fusion with chromatin-modulating peptides (CMPs), derived from high mobility group proteins HMGN1 and HMGB1, histone H1, and chromatin remodeling complexes, improves its activity by up to several fold, particularly on refractory target sites.