null allele mouse lines  (Charles River Laboratories)

Journal: BMC Biology

Article Title: Comparative analysis of single-stranded DNA donors to generate conditional null mouse alleles

doi: 10.1186/s12915-018-0529-0

Figure Lengend Snippet: Conditional KO ssODN ( a ) and lssDNA ( b ) targeting attempts. Each donut chart represents the summation of each allele type for all F0 mice genotyped, by ODN donor. In the center of the chart is the total number of F0 mice genotyped. Percentages of each allele type of the total number of mice genotyped are listed on each segment. 5′ and 3′ loxP: Includes animals genotyped for both 5′ and 3′ loxP sites, irrespective of the presence of any additional alleles (e.g., animals with 5′ loxP, 3′ loxP and a null allele detected); Null allele: Includes animals genotyped for a null allele, which may also have a single HDR and/or NHEJ indel event; Single HDR event: Includes animals genotyped for a single HDR event with or without additional indel events; NHEJ Indel event: Animals in which only indel alleles were observed. a Summary of genotypes identified from ssODN targeting attempts, and symmetric and asymmetric homology arm designs. b Summary of genotypes identified from four lssDNA targeting attempts

Article Snippet: The International Mouse Phenotyping Consortium (IMPC) and its NIH-funded Knockout Mouse Production and Phenotyping (KOMP 2 ) component is producing a null allele mouse line for every protein coding gene in the mouse genome, and phenotyping each null allele line to elucidate gene function in human biology and disease [ ].

Techniques:

Journal: BMC Biology

Article Title: Comparative analysis of single-stranded DNA donors to generate conditional null mouse alleles

doi: 10.1186/s12915-018-0529-0

Figure Lengend Snippet: Pearson correlations for 5′ and 3′ loxP sites. Data plotted based on number of F0 mice with an HDR event (x-axis) versus any evidence for a DSB generated at the respective sgRNA site, 5′ and 3′, such as a NHEJ indel, HDR event, or the formation of a null allele (y-axis)

Article Snippet: The International Mouse Phenotyping Consortium (IMPC) and its NIH-funded Knockout Mouse Production and Phenotyping (KOMP 2 ) component is producing a null allele mouse line for every protein coding gene in the mouse genome, and phenotyping each null allele line to elucidate gene function in human biology and disease [ ].

Techniques: Generated

Journal: BMC Biology

Article Title: Comparative analysis of single-stranded DNA donors to generate conditional null mouse alleles

doi: 10.1186/s12915-018-0529-0

Figure Lengend Snippet: a Copy number data from quantitative SYBR Green PCR assays for Abat , Kctd7 , Slc2a12 , and Uqcr10 , using primers following the design introduced in Fig. for both 5′ and 3′ loxP ssODN donors. All mice screened for each conditional null attempt were heterozygous conditional null mice from the N1 generation that were sequence confirmed for the conditional allele. Negative control samples were CRISPR-targeted N1 mice that were wild-type for the allele being analyzed. b, c Copy number data from TaqMan ® Copy Number assays for ( b ) Slc2a12 and Smc1a (paired ssODN conditional null targeting attempts) and ( c ) Eif2s2 and Cd44 (single lssDNA conditional null targeting attempts) using N1 progeny from a single founder. Genotypes are listed with the mouse ID. Animals with an asterisk to the right of the genotype were sequence-confirmed for the conditional null allele. Of note, Smc1a is located on the X chromosome, thereby males only have one copy of the gene. As in ( a ), negative control samples were CRISPR-targeted N1 mice that were wild-type for the allele being analyzed

Article Snippet: The International Mouse Phenotyping Consortium (IMPC) and its NIH-funded Knockout Mouse Production and Phenotyping (KOMP 2 ) component is producing a null allele mouse line for every protein coding gene in the mouse genome, and phenotyping each null allele line to elucidate gene function in human biology and disease [ ].

Techniques: SYBR Green Assay, Sequencing, Negative Control, CRISPR