... in PMC for immediate release. Data and resource availability The raw and processed single-cell RNA-sequencing datasets are available in the Gene Expression Omnibus (GEO) database under accession number GSE275013 . All mutant alleles were generated with the CRISPR/Cas9 gene-editing...

... involvement of the super elongation complex component MLLT3 in caudal neural tissue specification. Neural induction Chick NMPs CRISPR/cas9 Super elongation complex In vivo screen Francis Crick Institute http://dx.doi.org/10.13039/100010438 European Research Council http...

... of Gb-abd-A ( Fig. 5A ). Table 2. Efficiency of CRISPR/Cas9-mediated genome editing in G. bimaculatus Figure Number of eggs injected Number of injected embryos with eGFP expression or phenotype (% of injected eggs) Number of embryos developed by 7 d AEL (% of injected...

... analyses revealed differential gene expression between ybx1 −/− and control ovaries, with significantly increased expression of cdkn1a (p21), a cell cycle inhibitor, in ybx1 −/− follicles. While cdkn1a knockout via CRISPR/Cas9 was embryonically lethal, the heterozygote ( cdkn1a +/− ) displayed advanced...

... CRISPR/Cas9 genome editing in multiple species. CRISPR/Cas9 Oocyte delivery Vitellogenesis Gene editing Transgenesis National Institutes of Health R00HD099315 GM134577 Hibbitt Foundation National Institutes of Health The Eugene Bell Center...

... null mutant. The medaka mitfa ex6del1 and mitfb ex6del7 mutation was generated by CRISPR/Cas9 on the Nagoya background. In this work, the homozygous mutants are designated as mitfa −/− and mitfb −/− . The AB strain of the zebrafish Danio rerio was used as the WT. Zebrafish pax3a...

... by treatment with 2 μM progesterone. Matured oocytes were colored with vital dyes, transferred to egg-laying host females, recovered and fertilised as previously described ( Heasman et al., 1991 ; Houston, 2018 , 2019 ). Mutations in wnt11b.L were generated in F0 embryos using CRISPR/Cas9 RNP...

... currently used sea urchin. We leveraged this advantage to generate a knockout mutant of the sea urchin homolog of the drug transporter ABCB1, a major player in xenobiotic disposition for all animals. Using CRISPR/Cas9, we generated large fragment deletions of ABCB1 and used these readily detected deletions...

... of the motor domain acts as a STOP. In contrast, uncoordinated mitochondrial motility was observed in CRISPR/Cas9-GSK3β non-phosphorylatable-KHCS314A Drosophila larval axons, owing to decreased kinesin 1 attachment to microtubules and/or membranes, and reduced ATPase activity. Together, we propose that GSK3β...

..., the targeting strategy using the PGK-neo cassette affected the expression of the neighbor gene Mesp1 ( Takahashi et al., 2007 ). In this study, we re-established Mesp1 and Mesp2 single or Mesp1/Mesp2 double-null mutant mice without inserting the PGK-neo cassette using CRISPR/Cas9-mediated genome editing...

... size due to loss of barley LAXATUM.A / BLADE-ON-PETIOLE2 ( HvBOP2 ) gene function. Taken together, we reveal novel pleiotropic roles and regulatory interactions for an AP2 -like gene controlling floret and grain development in a temperate cereal. CRISPR/Cas9 technology was used to generate mutations...

... in our analyses, we used traditional conditional knockout approaches to validate the phenotype of OHC death following Rbm24 mutation. First, by using the CRISPR/Cas9 approach, we generated two Rbm24 mutant strains: (1) germline Rbm24 +/− knockout mice ( Fig. S4A-C ); and (2) conditional Rbm24...

...Masanori Abe; Reiko Kuroda ABSTRACT The establishment of left-right body asymmetry is a key biological process that is tightly regulated genetically. In the first application of CRISPR/Cas9 to a mollusc, we show decisively that the actin-related diaphanous gene Lsdia1 is the single maternal gene...

... to clarify their roles in sex reversal, biallelic mutants for crhr1 and crhr2 were produced by CRISPR/Cas9 technology. Remarkably, biallelic mutants of both loci ( crhr1 and crhr2 ) did not undergo female-to-male sex reversal upon exposure to HT. Inhibition of this process in double corticotropin-releasing...

... interaction and the compensatory mechanisms involved can make findings from single mutations, at best difficult to interpret, and, at worst, misleading. Hence, it is necessary to establish an efficient way to disrupt multiple genes simultaneously. CRISPR/Cas9-mediated base editing disrupts gene function...

...-directed repair (HDR) that provides efficient non-mosaic targeted insertion of small DNA fragments (40-50 nucleotides) in 4.4-25.7% of F0 tadpoles, with germline transmission. For both CRISPR/Cas9-mediated HDR gene editing and indel mutation, the gene-edited F0 embryos are uniformly heterozygous...

... for help with RB-KO human ESC design and development, and teratoma studies. Author contributions T.M., V.N.-E. and J.H. designed experiments and wrote the manuscript. T.M., V.N.-E. and S.K. performed and analyzed experiments. S.K. and J.W.S. generated the RB1 -KO human ESCs using CRISPR/Cas9...

... compatible with targeted genetic manipulations. However, gene targeting, which is crucial for functional studies of embryonic stem cells, has not been exploited to date in NSC lines. Here, we deploy CRISPR/Cas9 technology to demonstrate a variety of sophisticated genetic modifications via gene targeting...

... these limitations by developing single-step optimized inducible gene knockdown or knockout (sOPTiKD or sOPTiKO) platforms. These are based on genetic engineering of human genomic safe harbors combined with an improved tetracycline-inducible system and CRISPR/Cas9 technology. We exemplify the efficacy...

...Ira L. Blitz; Margaret B. Fish; Ken W. Y. Cho CRISPR/Cas9 genome editing is revolutionizing genetic loss-of-function analysis but technical limitations remain that slow progress when creating mutant lines. First, in conventional genetic breeding schemes, mosaic founder animals carrying mutant...