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i GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases
... observation using a dis- secting microscope (SZ11; Olympus, Tokyo, Japan), as described previously [10, 24] with slight ...ampulla using a ...(pulled using a P-97/IVF electric puller; Sutter ... See full document
15
Efficient homologous recombination mediated genome engineering in zebrafish using TALE nucleases
... custom nucleases has enabled new methods for disruption and editing of specific genes; however, the low efficiency of current HR-mediated genome editing methods has limited their utility in zebrafish (Zu et ... See full document
12
Perfectly matched 20 nucleotide guide RNA sequences enable robust genome editing using high fidelity SpCas9 nucleases
... The CRISPR-Cas9 system recognizes genomic sites via Watson-Crick base pairing by virtue of 20-nucleotide (nt) guide sequences in the guide RNAs (gRNAs) that direct Cas9 for targeted cleavage [1 – ...The ... See full document
7
Run on of germline apoptosis promotes gonad senescence in C elegans
... occur, gonad disintegration does not occur ...male gonad did not cause senescent atrophy (Supplementary Figure 5), suggesting that absence of major atrophy in wild-type males is not simply attributable to ... See full document
16
Run-on of germline apoptosis promotes gonad senescence in C. elegans
... distal gonad degeneration and ...age-related gonad degeneration. We document the continuation of germline apoptosis in later life, and report that genetically blocking or increasing PA retards or ... See full document
15
CRISPR Genome Engineering for Human Pluripotent Stem Cell Research
... in CRISPR-Cas9 systems ...of CRISPR way beyond loss-of-function experiments ...such method, CASFISH [165], which is Cas9-mediated fluorescence in situ hybridization, is a rapid cost-effective ... See full document
25
The control of sexual identity in the Drosophila germline
... Interestingly, the Jak/Stat pathway is activated in all germ cells as they enter the male embryonic gonad, but is then active only in GSCs in the adult testis. Thus, does the Jak/Stat pathway promote male germ ... See full document
9
Efficient Editing of Malaria Parasite Genome Using the CRISPR/Cas9 System
... Cas9/sgRNA method has other advantages compared with the traditional gene editing ...conventional method of gene modification based on HR generally requires insertion of selection markers and other plasmid ... See full document
9
What is the available evidence for the range of applications of genome-editing as a new tool for plant trait modification and the potential occurrence of associated off-target effects: a systematic map
... decades, genome-editing techniques such as CRISPR/Cas, TALENs, Zinc-Finger Nucle- ases, Meganucleases, Oligonucleotide-Directed Mutagenesis and base editing have been developed enabling a precise ... See full document
33
CRISPR-Based Methods for Caenorhabditis elegans Genome Engineering
... the genome in an error-free ...the genome, and longer homology arms led to reduced efficiency (Paix et ...obtained using 500- to 700-bp homology arms, but occurred rarely or not at all when ... See full document
17
Genome Editing with Engineered Nucleases in Economically Important Animals and Plants: State of the Art in the Research Pipeline
... the genome of an acceptor organism to obtain a desired phenotype (such as disease- or pathogen- resistance in crops, and meat production or decreased allergenicity in animals) thus forming so-called genetically ... See full document
22
Optimizing a CRISPR-Cpf1-based genome engineering system for Corynebacterium glutamicum
... pYJ1 using T4 DNA ...ATCC13032 genome was used as a template to amplify 500 bp upstream and down- stream homologous arms using corresponding prim- ers (such as F5/R5 and F6/R6, ...cut using ... See full document
13
CRISPR-Cas12a (Cpf1): a versatile tool in the plant genome editing tool box for agricultural advancement
... innovative method to increase HDR that has been applied in yeast systems is CRISPEY (Sharon et ...other genome editing ...plant genome editing tool for crop improvement (Lin et ... See full document
27
High efficiency non mosaic CRISPR mediated knock in and indel mutation in F0 Xenopus
... maternal genome was mutated because Cas9 protein and/or the sgRNA was degraded over the 72-h culture ...paternal genome is likely to occur in embryos generated from these ...that CRISPR ... See full document
7
Cutting it Right: Plasticity and Strategy of CRISPR RNA Specific Nucleases
... subtypes I-A, I-B, I-D, III-A and III- B has been characterized in ...the CRISPR repeat, anchoring it in position for the cleavage at 3’-end of the repeat on the opposite surface of the ... See full document
23
Advances in Engineering the Fly Genome with the CRISPR-Cas System
... all CRISPR experiments is a gRNA for targeting Cas9 to a specific ...the genome that might be subject to off-target ...a genome will define the number and position of target ...the genome of the ... See full document
18
Keeping CRISPR/Cas on-Target
... for genome targeting purposes(Ran et ...of CRISPR/Cas system (Cong et ...based CRISPR interference (CRISPRi) is proven to be more significant than classic RNAi-based ...2013). CRISPR/ Cas ... See full document
13
Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9
... locus using CRISPR/Cas9. (A) Strategy for constructing the CRISPR/Cas9 plasmid pU6-SAG1: chiRNA encoding a protospacer specific to SAG1 (blue) and the Cas9 recognition motif (orange) was cloned under ... See full document
8
Precise and Heritable Genome Editing in Evolutionarily Diverse Nematodes Using TALENs and CRISPR/Cas9 to Engineer Insertions and Deletions
... custom-designed nucleases to induce DNA double-strand breaks (DSBs) at genomic locations of choice has transformed our ability to edit genomes, regardless of their ...strategies using site-specific ... See full document
26
CrEdit: CRISPR mediated multi-loci gene integration in Saccharomyces cerevisiae
... for genome integra- tions using selection ...recombination using 500 bp long homology arms, this method results in successful integration into a single site ...loci using selection ... See full document
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