Matches in SemOpenAlex for { <https://semopenalex.org/work/W3196301293> ?p ?o ?g. }
- W3196301293 endingPage "9429" @default.
- W3196301293 startingPage "9429" @default.
- W3196301293 abstract "CRISPR-Cas9-based genome-editing is a highly efficient and cost-effective method to generate zebrafish loss-of-function alleles. However, introducing patient-specific variants into the zebrafish genome with CRISPR-Cas9 remains challenging. Targeting options can be limited by the predetermined genetic context, and the efficiency of the homology-directed DNA repair pathway is relatively low. Here, we illustrate our efficient approach to develop knock-in zebrafish models using two previously variants associated with hereditary sensory deficits. We employ sgRNA-Cas9 ribonucleoprotein (RNP) complexes that are micro-injected into the first cell of fertilized zebrafish eggs together with an asymmetric, single-stranded DNA template containing the variant of interest. The introduction of knock-in events was confirmed by massive parallel sequencing of genomic DNA extracted from a pool of injected embryos. Simultaneous morpholino-induced blocking of a key component of the non-homologous end joining DNA repair pathway, Ku70, improved the knock-in efficiency for one of the targets. Our use of RNP complexes provides an improved knock-in efficiency as compared to previously published studies. Correct knock-in events were identified in 3-8% of alleles, and 30-45% of injected animals had the target variant in their germline. The detailed technical and procedural insights described here provide a valuable framework for the efficient development of knock-in zebrafish models." @default.
- W3196301293 created "2021-09-13" @default.
- W3196301293 creator A5000563716 @default.
- W3196301293 creator A5004308908 @default.
- W3196301293 creator A5010528169 @default.
- W3196301293 creator A5014529631 @default.
- W3196301293 creator A5020260568 @default.
- W3196301293 creator A5022569376 @default.
- W3196301293 creator A5044496374 @default.
- W3196301293 creator A5089656819 @default.
- W3196301293 date "2021-08-30" @default.
- W3196301293 modified "2023-09-30" @default.
- W3196301293 title "Efficient Generation of Knock-In Zebrafish Models for Inherited Disorders Using CRISPR-Cas9 Ribonucleoprotein Complexes" @default.
- W3196301293 cites W1864853872 @default.
- W3196301293 cites W1977709885 @default.
- W3196301293 cites W1984456138 @default.
- W3196301293 cites W2052635096 @default.
- W3196301293 cites W2053454212 @default.
- W3196301293 cites W2060297895 @default.
- W3196301293 cites W2074416973 @default.
- W3196301293 cites W2093767301 @default.
- W3196301293 cites W2124715997 @default.
- W3196301293 cites W2127497550 @default.
- W3196301293 cites W2145666712 @default.
- W3196301293 cites W2164695230 @default.
- W3196301293 cites W2260480416 @default.
- W3196301293 cites W2289992440 @default.
- W3196301293 cites W2343277111 @default.
- W3196301293 cites W2343876552 @default.
- W3196301293 cites W2550769902 @default.
- W3196301293 cites W2793309516 @default.
- W3196301293 cites W2803498661 @default.
- W3196301293 cites W2807817820 @default.
- W3196301293 cites W2889830609 @default.
- W3196301293 cites W2894792758 @default.
- W3196301293 cites W2896237399 @default.
- W3196301293 cites W2897726150 @default.
- W3196301293 cites W2910850705 @default.
- W3196301293 cites W3006244701 @default.
- W3196301293 cites W3008149374 @default.
- W3196301293 cites W3026324878 @default.
- W3196301293 cites W3041014829 @default.
- W3196301293 cites W3109270346 @default.
- W3196301293 cites W3158544805 @default.
- W3196301293 cites W3196085166 @default.
- W3196301293 doi "https://doi.org/10.3390/ijms22179429" @default.
- W3196301293 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/8431507" @default.
- W3196301293 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/34502338" @default.
- W3196301293 hasPublicationYear "2021" @default.
- W3196301293 type Work @default.
- W3196301293 sameAs 3196301293 @default.
- W3196301293 citedByCount "9" @default.
- W3196301293 countsByYear W31963012932022 @default.
- W3196301293 countsByYear W31963012932023 @default.
- W3196301293 crossrefType "journal-article" @default.
- W3196301293 hasAuthorship W3196301293A5000563716 @default.
- W3196301293 hasAuthorship W3196301293A5004308908 @default.
- W3196301293 hasAuthorship W3196301293A5010528169 @default.
- W3196301293 hasAuthorship W3196301293A5014529631 @default.
- W3196301293 hasAuthorship W3196301293A5020260568 @default.
- W3196301293 hasAuthorship W3196301293A5022569376 @default.
- W3196301293 hasAuthorship W3196301293A5044496374 @default.
- W3196301293 hasAuthorship W3196301293A5089656819 @default.
- W3196301293 hasBestOaLocation W31963012931 @default.
- W3196301293 hasConcept C104317684 @default.
- W3196301293 hasConcept C132455925 @default.
- W3196301293 hasConcept C144501496 @default.
- W3196301293 hasConcept C2776878037 @default.
- W3196301293 hasConcept C2796436 @default.
- W3196301293 hasConcept C54355233 @default.
- W3196301293 hasConcept C67705224 @default.
- W3196301293 hasConcept C70721500 @default.
- W3196301293 hasConcept C75934600 @default.
- W3196301293 hasConcept C86803240 @default.
- W3196301293 hasConcept C98108389 @default.
- W3196301293 hasConceptScore W3196301293C104317684 @default.
- W3196301293 hasConceptScore W3196301293C132455925 @default.
- W3196301293 hasConceptScore W3196301293C144501496 @default.
- W3196301293 hasConceptScore W3196301293C2776878037 @default.
- W3196301293 hasConceptScore W3196301293C2796436 @default.
- W3196301293 hasConceptScore W3196301293C54355233 @default.
- W3196301293 hasConceptScore W3196301293C67705224 @default.
- W3196301293 hasConceptScore W3196301293C70721500 @default.
- W3196301293 hasConceptScore W3196301293C75934600 @default.
- W3196301293 hasConceptScore W3196301293C86803240 @default.
- W3196301293 hasConceptScore W3196301293C98108389 @default.
- W3196301293 hasFunder F4320309607 @default.
- W3196301293 hasFunder F4320325929 @default.
- W3196301293 hasIssue "17" @default.
- W3196301293 hasLocation W31963012931 @default.
- W3196301293 hasLocation W31963012932 @default.
- W3196301293 hasLocation W31963012933 @default.
- W3196301293 hasLocation W31963012934 @default.
- W3196301293 hasLocation W31963012935 @default.
- W3196301293 hasOpenAccess W3196301293 @default.
- W3196301293 hasPrimaryLocation W31963012931 @default.
- W3196301293 hasRelatedWork W2180658822 @default.
- W3196301293 hasRelatedWork W2546906832 @default.