Ervin Welker
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View article: A cleavage rule for selection of increased-fidelity SpCas9 variants with high efficiency and no detectable off-targets
A cleavage rule for selection of increased-fidelity SpCas9 variants with high efficiency and no detectable off-targets Open
View article: Position-dependent sequence motif preferences of SpCas9 are largely determined by scaffold-complementary spacer motifs
Position-dependent sequence motif preferences of SpCas9 are largely determined by scaffold-complementary spacer motifs Open
Streptococcus pyogenes Cas9 (SpCas9) nuclease exhibits considerable position-dependent sequence preferences. The reason behind these preferences is not well understood and is difficult to rationalise, since the protein establishes interact…
View article: SuperFi-Cas9 exhibits remarkable fidelity but severely reduced activity yet works effectively with ABE8e
SuperFi-Cas9 exhibits remarkable fidelity but severely reduced activity yet works effectively with ABE8e Open
View article: Target-matched increased fidelity SpCas9 variants can edit any target without genome-wide off-target effects
Target-matched increased fidelity SpCas9 variants can edit any target without genome-wide off-target effects Open
Streptococcus pyogenes Cas9 (SpCas9) has been employed as a genome engineering tool with a promising potential within therapeutics. However, its off- target effects pose a major limitation for applications requiring high specificity. Appro…
View article: SuperFi-Cas9 exhibits extremely high fidelity but reduced activity in mammalian cells
SuperFi-Cas9 exhibits extremely high fidelity but reduced activity in mammalian cells Open
Several advances have been made to SpCas9, the most widely used CRISPR/Cas genome editing tool, to reduce its unwanted off-target effects. The most promising approach is the development of increased fidelity nuclease (IFN) variants of SpCa…
View article: PEAR, a flexible fluorescent reporter for the identification and enrichment of successfully prime edited cells
PEAR, a flexible fluorescent reporter for the identification and enrichment of successfully prime edited cells Open
Prime editing is a recently developed CRISPR/Cas9 based gene engineering tool that allows the introduction of short insertions, deletions, and substitutions into the genome. However, the efficiency of prime editing, which typically achieve…
View article: Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin
Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin Open
The cellular prion protein (PrPC) is renowned for its infectious conformational isoform PrPSc, capable of templating subsequent conversions of healthy PrPCs and thus triggering the group of incurable diseases known as transmissible spongif…
View article: Author response: PEAR, a flexible fluorescent reporter for the identification and enrichment of successfully prime edited cells
Author response: PEAR, a flexible fluorescent reporter for the identification and enrichment of successfully prime edited cells Open
View article: BEAR reveals that increased fidelity variants can successfully reduce the mismatch tolerance of adenine but not cytosine base editors
BEAR reveals that increased fidelity variants can successfully reduce the mismatch tolerance of adenine but not cytosine base editors Open
View article: PEAR: a flexible fluorescent reporter for the identification and enrichment of successfully prime edited cells
PEAR: a flexible fluorescent reporter for the identification and enrichment of successfully prime edited cells Open
Prime editing is a recently developed gene engineering tool that allows the introduction of short insertions, deletions or substitutions into the genome. However, the efficiency of prime editing, generally reaching around 10-30% editing, h…
View article: The G127V variant of the prion protein interferes with dimer formation in vitro but not in cellulo
The G127V variant of the prion protein interferes with dimer formation in vitro but not in cellulo Open
View article: A method for characterizing Cas9 variants via a one-million target sequence library of self-targeting sgRNAs
A method for characterizing Cas9 variants via a one-million target sequence library of self-targeting sgRNAs Open
Detailed target-selectivity information and experiment-based efficacy prediction tools are primarily available for Streptococcus pyogenes Cas9 (SpCas9). One obstacle to develop such tools is the rarity of accurate data. Here, we report a m…
View article: BEAR reveals that increased fidelity variants can successfully reduce the mismatch-tolerance of adenine but not cytosine base editors
BEAR reveals that increased fidelity variants can successfully reduce the mismatch-tolerance of adenine but not cytosine base editors Open
Adenine and cytosine base editors (ABE, CBE) are designed to generate single base mutations in genes without necessarily generating DNA double-strand breaks and undesired indel mutations. However, the activity of base editors employing an …
View article: Blackjack mutations improve the on-target activities of increased fidelity variants of SpCas9 with 5′G-extended sgRNAs
Blackjack mutations improve the on-target activities of increased fidelity variants of SpCas9 with 5′G-extended sgRNAs Open
View article: Improved LbCas12a variants with altered PAM specificities further broaden the genome targeting range of Cas12a nucleases
Improved LbCas12a variants with altered PAM specificities further broaden the genome targeting range of Cas12a nucleases Open
The widespread use of Cas12a (formerly Cpf1) nucleases for genome engineering is limited by their requirement for a rather long TTTV protospacer adjacent motif (PAM) sequence. Here we have aimed to loosen these PAM constraints and have gen…
View article: Mb- and FnCpf1 nucleases are active in mammalian cells: activities and PAM preferences of four wild-type Cpf1 nucleases and of their altered PAM specificity variants
Mb- and FnCpf1 nucleases are active in mammalian cells: activities and PAM preferences of four wild-type Cpf1 nucleases and of their altered PAM specificity variants Open
Cpf1s, the RNA-guided nucleases of the class II clustered regularly interspaced short palindromic repeats system require a short motive called protospacer adjacent motif (PAM) to be present next to the targeted sequence for their activity.…
View article: Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage
Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage Open
View article: Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage
Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage Open
Background The propensity for off-target activity of Streptococcus pyogenes Cas9 (SpCas9) has been considerably decreased by rationally engineered variants with increased fidelity (eSpCas9; SpCas9-HF1). However, a subset of targets still g…
View article: A convenient method to pre-screen candidate guide RNAs for CRISPR/Cas9 gene editing by NHEJ-mediated integration of a ‘self-cleaving’ GFP-expression plasmid
A convenient method to pre-screen candidate guide RNAs for CRISPR/Cas9 gene editing by NHEJ-mediated integration of a ‘self-cleaving’ GFP-expression plasmid Open
The efficacies of guide RNAs (gRNAs), the short RNA molecules \nthat bind to and determine the sequence specificity of the \nStreptococcus pyogenes Cas9 nuclease, to mediate DNA cleavage \nvary dramatically. Thus, the selection of appropri…
View article: The small molecule AUTEN-99 (autophagy enhancer-99) prevents the progression of neurodegenerative symptoms
The small molecule AUTEN-99 (autophagy enhancer-99) prevents the progression of neurodegenerative symptoms Open
View article: Additional file 4: of Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage
Additional file 4: of Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage Open
Statistics. (XLSX 54 kb)
View article: Additional file 2: of Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage
Additional file 2: of Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage Open
Primer, sequence, and deep sequencing data. (XLSX 36 kb)
View article: The prion protein family member Shadoo induces spontaneous ionic currents in cultured cells
The prion protein family member Shadoo induces spontaneous ionic currents in cultured cells Open
View article: Cpf1 nucleases demonstrate robust activity to induce DNA modification by exploiting homology directed repair pathways in mammalian cells
Cpf1 nucleases demonstrate robust activity to induce DNA modification by exploiting homology directed repair pathways in mammalian cells Open
View article: Expression of the Prion Protein Family Member Shadoo Causes Drug Hypersensitivity That Is Diminished by the Coexpression of the Wild Type Prion Protein
Expression of the Prion Protein Family Member Shadoo Causes Drug Hypersensitivity That Is Diminished by the Coexpression of the Wild Type Prion Protein Open
View article: Prion Protein Does Not Confer Resistance to Hippocampus-Derived Zpl Cells against the Toxic Effects of Cu2+, Mn2+, Zn2+ and Co2+ Not Supporting a General Protective Role for PrP in Transition Metal Induced Toxicity
Prion Protein Does Not Confer Resistance to Hippocampus-Derived Zpl Cells against the Toxic Effects of Cu2+, Mn2+, Zn2+ and Co2+ Not Supporting a General Protective Role for PrP in Transition Metal Induced Toxicity Open
The interactions of transition metals with the prion protein (PrP) are well-documented and characterized, however, there is no consensus on their role in either the physiology of PrP or PrP-related neurodegenerative disorders. PrP has been…