pac sgrna cas9 targeting  (Addgene inc)

Journal: Scientific Reports

Article Title: Genome editing of WSSV CRISPR/Cas9 and immune activation extends the survival of infected Penaeus vannamei

doi: 10.1038/s41598-024-78277-7

Figure Lengend Snippet: Analysis of gRNAs and Cas9 expression induction from CRISPR/Cas9-WSSV delivery in hemocytes and P. vannamei tissue via RT‒PCR. ( a ) Amplicons of RT‒PCR detection of gRNA1, gRNA2 and Cas from CRISPR/Cas9-WSSV plasmid-transfected cultured hemocytes at 12 h and 24 h. The amplification of EF-1α was used as an internal control, and pAC-sgRNA-transfected cells were used as a negative control (C). ( b ) Analysis of gRNAs and Cas9 expression in shrimp tissues.

Article Snippet: The chimeric DNA was double digested with EcoRI and KpnI and inserted into the pAC-sgRNA-Cas9 vector (Addgene, USA).

Techniques: Expressing, CRISPR, Plasmid Preparation, Transfection, Cell Culture, Amplification, Control, Negative Control

Journal: Scientific Reports

Article Title: Genome editing of WSSV CRISPR/Cas9 and immune activation extends the survival of infected Penaeus vannamei

doi: 10.1038/s41598-024-78277-7

Figure Lengend Snippet: Color inverted picture of the agarose gel electrophoresis of cleaved target DNA by the CRISPR/Cas9-WSSV reaction. Lane M: DNA markers; Lane 1: reaction of the WSSV143 amplicon and gRNA specific to GFP and Caspase 9 (negative control); Lane 2: reaction of the WSSV 143 amplicon with gRNA1 and Cas9; Lane 3: reaction of WSSV360, gRNA2 and Cas9; Lane 4: reaction of the IHHNV amplicon and gRNA specific to IHHNV 9397 and Cas9 (positive control).

Article Snippet: The chimeric DNA was double digested with EcoRI and KpnI and inserted into the pAC-sgRNA-Cas9 vector (Addgene, USA).

Techniques: Agarose Gel Electrophoresis, CRISPR, Amplification, Negative Control, Positive Control

Journal: Scientific Reports

Article Title: Genome editing of WSSV CRISPR/Cas9 and immune activation extends the survival of infected Penaeus vannamei

doi: 10.1038/s41598-024-78277-7

Figure Lengend Snippet: CRISPR/Cas9 WSSV was introduced into the shrimp genome to suppress viral replication and rescue shrimp. WSSV was detected by qRT‒PCR at 24-72 h. WSSV DNA expression in the gills of infected P. vannamei under different treatments and at different time points.

Article Snippet: The chimeric DNA was double digested with EcoRI and KpnI and inserted into the pAC-sgRNA-Cas9 vector (Addgene, USA).

Techniques: CRISPR, Expressing, Infection

Journal: Scientific Reports

Article Title: Genome editing of WSSV CRISPR/Cas9 and immune activation extends the survival of infected Penaeus vannamei

doi: 10.1038/s41598-024-78277-7

Figure Lengend Snippet: Analysis of immune-related gene expression in P. vannamei in response to WSSV challenge and CRISPR/Cas9-WSSV treatment. qRT‒PCR determination of HSP70 expression in the hepatopancreas ( a ) and muscle ( b ) and the expression of the lectin gene in the hepatopancreas ( c ) and muscle (d) at different times post infection. The data are presented as mean ± SD ( n = 3). The asterisk indicates significant differences between groups at each time point (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The chimeric DNA was double digested with EcoRI and KpnI and inserted into the pAC-sgRNA-Cas9 vector (Addgene, USA).

Techniques: Expressing, CRISPR, Infection

Journal: Scientific Reports

Article Title: Genome editing of WSSV CRISPR/Cas9 and immune activation extends the survival of infected Penaeus vannamei

doi: 10.1038/s41598-024-78277-7

Figure Lengend Snippet: Survival rate of P. vannamei infected with 1 × 10 3 Copies/100 µL of WSSV or subjected to CRISPR/Cas9-WSSV treatment.

Article Snippet: The chimeric DNA was double digested with EcoRI and KpnI and inserted into the pAC-sgRNA-Cas9 vector (Addgene, USA).

Techniques: Infection, CRISPR