CRISPR/Cas9‐mediated mstnb1 disruption enhances growth in rainbow trout

Programmable RNA targeting with clustered regularly interspaced short palindromic repeats (CRISPR) effector Cas7-11 in zebrafish embryos and mammalian cells

Findings establish Cas7-11 as a precise and efficient RNA knockdown tool for functional studies in embryonic development and stem cell biology, providing a versatile alternative to DNA-based gene-editing approaches.

CRISPR/Cas12a-mediated allele engineering of SmAPRR2 and SmGLK2 reveals complementary control of fruit peel and flesh chlorophyll pigmentation in eggplant

A multiplex CRISPR/Cas12a system is established in eggplant accession MEL3, representing, to the authors' knowledge, the first application of this nuclease for genome editing in eggplant and demonstrating the potential of Cas12a for functional genomics, allele engineering, and precision breeding in eggplant.

Homologous knock-in processes facilitated by CRISPR-Cas9 in Aurantiochytrium and disrupting the gene for DNA ligase IV prevents transformation events

This study explores advanced molecular breeding techniques for Aurantiochytrium limacinum , a eukaryotic microorganism valued for its industrial production of DHA and astaxanthin. We first employed the CRISPR-Cas9 system to simultaneously disrupt two genes: crtIBY , a multifunctional carotenoid synthesis gene used as a visual marker, and lig4 , which is involved in non-homologous end joining (NHEJ). This genetic disruption caused the wild-type orange colonies to turn into a white colony (Δ lig4 -Δ crtIBY -TA#6), indicating the loss of carotenoid production. Subsequently, we successfully demonstrated marker recycling by repairing the crtIBY gene using single-strand oligodeoxynucleotide (ssODN) templates, which restored the orange phenotype (Δ lig4 -TB#18). Further investigation into the Δ lig4 strain (TB#18) revealed that its overall transformation efficiency dropped significantly compared to that of the wild-type strain when using only a bleomycin resistance expression cassette with two crtIBY homologous regions. However, when we combined the bleomycin resistance expression cassette with CRISPR-Cas9 ribonucleoproteins (RNPs) during electroporation, the Δ lig4 strains (TA#6 and TB#18) exhibited higher homologous recombination-type efficiencies through double-crossover-type events than the wild-type. These results indicate that while Lig4 is important for general transformation against zeocin, disrupting the NHEJ pathway enhances precise gene targeting. This study demonstrated the potential of these CRISPR-Cas9 systems to serve as a robust foundation for the functional genomics and metabolic engineering of A. limacinum .

CRISPR/Cas9-Mediated Generation and Characterization of an Ent2*/CyO Drosophila melanogaster Strain.

The phenotypic profile of a CRISPR-generated Ent2 mutant line is described and the feasibility of combining genome editing with balancer chromosome strategies in Drosophila is demonstrated, demonstrating the feasibility of combining genome editing with balancer chromosome strategies in Drosophila.

Ef fi cient epigenetic editing of the zebra fi sh slc45a2 gene using the CRISPR/dCas9 system and tandem guide RNAs

Precise dual-gene knockout of MSTN and SOCS2 via cytidine base editing enhances muscling and growth in goats