CRISPR/Cas9-mediated genome editing of PML in human cell lines.

Slides:

Advertisements

Similar presentations

A b Fig. S1 Expression constructs for Cas9 without DsRed gene, and Cas9 mRNA level in pZD_Cas9 transformed calli. a pZH_Cas9 without the DsRed expression.

Advertisements

Molecular Therapy - Nucleic Acids

Molecular Therapy - Nucleic Acids

Targeted Disruption of V600E-Mutant BRAF Gene by CRISPR-Cpf1

Figure 1. AsCpf1 and LbCpf1-mediated gene editing in human cells

Figure S1. Analysis of the embryo edited using gRNA-2 and HF2-BE2

Yuming Lu, Jian-Kang Zhu Molecular Plant

Dan Ding, Kaiyuan Chen, Yuedan Chen, Hong Li, Kabin Xie

by Megan D. Hoban, Gregory J. Cost, Matthew C

by Fawwaz Yassin, Sheldon P. Rothenberg, Sreedhar Rao, Marilyn M

CRISPR/Cas9-Mediated Scanning for Regulatory Elements Required for HPRT1 Expression via Thousands of Large, Programmed Genomic Deletions Molly Gasperini,

Strategy for CRISPR/Cas9-mediated genome editing in ΔEx50 mice

Molecular Therapy - Nucleic Acids

Molecular Therapy - Methods & Clinical Development

Molecular Therapy - Nucleic Acids

Figure 1 Adaptive immune system of bacteria and archaea

In Vivo Knockout of the Vegfa Gene by Lentiviral Delivery of CRISPR/Cas9 in Mouse Retinal Pigment Epithelium Cells Andreas Holmgaard, Anne Louise Askou,

Osteoarthritis and Cartilage

Volume 13, Issue 6, Pages (December 2013)

USH2A Gene Editing Using the CRISPR System

Supplemental Figure 3 A B C T-DNA 1 2 RGLG1 2329bp 3 T-DNA 1 2 RGLG2

Molecular Therapy - Nucleic Acids

RNA-Guided Genome Editing in Plants Using a CRISPR–Cas System

Volume 15, Issue 5, Pages (November 2014)

Andrew R. Bassett, Charlotte Tibbit, Chris P. Ponting, Ji-Long Liu

Jung-Ok Han, Sharri B Steen, David B Roth Molecular Cell

Volume 153, Issue 4, Pages (May 2013)

Yubao Wang, Ben B. Li, Jing Li, Thomas M. Roberts, Jean J. Zhao

Chromatin Remodeling In Vivo

RAD51 is essential for L. donovani.

Gang Wang, Na Zhao, Ben Berkhout, Atze T Das Molecular Therapy

Volume 9, Issue 4, Pages (November 2014)

Targeted Myostatin Gene Editing in Multiple Mammalian Species Directed by a Single Pair of TALE Nucleases Li Xu, Piming Zhao, Andrew Mariano, Renzhi.

GFP to BFP Conversion: A Versatile Assay for the Quantification of CRISPR/Cas9- mediated Genome Editing Astrid Glaser, Bradley McColl, Jim Vadolas Molecular.

Volume 10, Issue 2, Pages (February 2018)

Volume 11, Issue 6, Pages (May 2015)

WRKY20 dual sgRNA approach.

Volume 25, Issue 11, Pages (November 2017)

Volume 25, Issue 1, Pages (January 2017)

Molecular Therapy - Methods & Clinical Development

William T. Hendriks, Curtis R. Warren, Chad A. Cowan Cell Stem Cell

Volume 153, Issue 4, Pages (May 2013)

Molecular Therapy - Nucleic Acids

Molecular Therapy - Nucleic Acids

Volume 23, Issue 3, Pages (March 2015)

CRISPR/Cas9-Mediated Deletion of CTG Expansions Recovers Normal Phenotype in Myogenic Cells Derived from Myotonic Dystrophy 1 Patients Claudia Provenzano,

Volume 17, Issue 2, Pages (August 2015)

Volume 26, Issue 6, Pages (June 2018)

Generation of heterozygous mutations with the CRISPR-Cas9 system.

Ciaran M Lee, Thomas J Cradick, Gang Bao Molecular Therapy

Volume 15, Issue 2, Pages (August 2014)

Inactivation of the Dp1 locus.

Gene editing by CRISPR/Cas9 for gene inactivation and targeted sequence replacement. Gene editing by CRISPR/Cas9 for gene inactivation and targeted sequence.

Volume 16, Issue 2, Pages (February 2015)

CRISPR/Cas9: Transcending the Reality of Genome Editing

Fig. 4 Gene disruption via chip.

Efficient In Vivo Liver-Directed Gene Editing Using CRISPR/Cas9

Multiplexed Mutagenesis Using the Csy4 System in Tomato Protoplasts.

Editing and reconstitution of CPAR2_

Molecular Therapy - Nucleic Acids

Volume 13, Issue 3, Pages (October 2015)

Targeted chromosomal translocations generated by targeting sites from two different chromosomes simultaneously. Targeted chromosomal translocations generated.

Molecular Therapy - Nucleic Acids

Molecular Therapy - Methods & Clinical Development

Volume 26, Issue 11, Pages (November 2018)

Volume 10, Issue 2, Pages (February 2018)

Genome-Edited Triple-Recessive Mutation Alters Seed Dormancy in Wheat

Knock-in of the rpl42-P56Q mutation using the split-ura4 system.

Fig. 3 Genome editing of the MSTN gene.

Presentation transcript:

CRISPR/Cas9-mediated genome editing of PML in human cell lines. CRISPR/Cas9-mediated genome editing of PML in human cell lines. (A) The Cas9 nuclease was targeted to exon 2 of the PML gene (green) by two selected gRNAs, whose binding sites are shown in blue (PAM motifs are in red). Arrows indicate the positions of the binding sites for the ODNs used in the PCR-Surveyor assay (blue arrows for gRNA1-guided and red arrows for gRNA2-guided cut sites). The results of the Surveyor assay are shown in the lower panel. Briefly, PCR products amplified from cells transfected with pLCv2-hPML1, pLCv2-hPML2, or pLCv2-CAG (Ctrl) were subjected to denaturation, reannealing, and digestion with the Surveyor enzyme. Arrowheads indicate cleavage products of the expected size. M, molecular size markers. (B) Sanger sequencing analysis of PML in cells transduced with LCv2-hPML1. THP-1 cells were transduced with lentiviral vectors produced using pLCv2-hPML1 or the control vector, pLCv2-CAG. Following puromycin selection, the targeted PML locus was PCR amplified and the PCR product was Sanger sequenced. The figure shows an alignment of the obtained sequence plots. (C) Decomposition of sequencing plots by TIDE assay. The graph shows the percentages of aberrant peaks upstream and downstream of the cut site in the sequencing reactions shown in panel B. The percentage of indel-containing alleles was computed by the TIDE assay. Nasser Masroori et al. mSphere 2017; doi:10.1128/mSphereDirect.00233-17