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Submissions Details

VALIDATION - The Jackson Laboratory Gene Editing Testing Center - Chen Validation (Chen, Zheng-Yi)
SCGE ID:1044 - Submission Date: 2021-04-01 
Experiment Name Type Description
Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear

In Vivo Delivery of CRISPR/Cas9 via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice
Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear

In Vivo Delivery of CRISPR/Cas9 editor via bioreducible lipid nanoparticle to the inner ear in Ai14 mice
VALIDATION - The Jackson Laboratory Gene Editing Testing Center - Gong Validation (Gong, Shaoqin)
SCGE ID:1046 - Submission Date: 2021-04-01 
Experiment Name Type Description
Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain

In Vivo Delivery of CRISPR/Cas9 via RNP-loaded nanocages to the brain in Ai14 mice
VALIDATION - The Jackson Laboratory Gene Editing Testing Center - Sontheimer Validation (Sontheimer, Eric)
SCGE ID:1048 - Submission Date: 2021-04-01 
Experiment Name Type Description
Independent validation of Sontheimer delivery platform using heavily modified guide RNAs complexed with Cas9 proteins to deliver CRISPR/Cas9 to mouse brain

In Vivo Heavily modified guide RNAs complexed with Cas9 proteins are injected locally to mouse striatum to activate reporter gene (mGFP). Editing detected via DAB staining in coronal brain sections.
NEW MODEL DEVELOPMENT - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC)
SCGE ID:1068 - Submission Date: 2022-08-11 
Experiment Name Type Description
Validating Traffic Light Reporter 2 (TLR2) Mouse Model in Homozygous Blastocysts using SpCas9

In Vitro Male and female homozygous mice mouse containing the Traffic Light Reporter 2 (TLR2) reporter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Cas9 RNP along with test guides and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for tagRFP fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 156-193. Total blast analyzed by Sanger sequencing, 84-120.
Validating Traffic Light Reporter 2 (TLR2) Mouse Model in Heterozygous Blastocysts using SaCas9

In Vitro WT female mouse and homozygous male mouse containing the Traffic Light Reporter 2 (TLR2) reporter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Cas9 RNP along with test guides and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for tagRFP fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced. ICE analysis tool from Synthego was used to score edits. NOTE: Total blasts analyzed expressing the fluorescent reporter, 55-67. Total blast analyzed by Sanger sequencing, 55-67.
Validating Traffic Light Reporter 2 (TLR2) Mouse Model in Heterozygous Blastocysts using SpCas9

In Vitro WT female mouse and homozygous male mouse containing the Traffic Light Reporter 2 (TLR2) reporter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Cas9 RNP along with test guides and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for tagRFP fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 46-120. Total blast analyzed by Sanger sequencing, 8-64.
NEW MODEL DEVELOPMENT - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - GER10 small animal reporter, submission 1
SCGE ID:1075 - Submission Date: 2022-10-19 
Experiment Name Type Description
Validating Gene Editing Reporter 10 (GER10) Mouse Model in Heterozygous Blastocysts

In Vitro WT female mouse and homozygous male mouse containing the Gene Editing Reporter 10 (GER10) reproter driven by the CAG promoter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Adenine Base Editor RNP complex and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for Venus (GFP) fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced to determine efficiency of base changes at the target region. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 174. Total blast analyzed by Sanger sequencing, 112.
NEW MODEL DEVELOPMENT - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - GER12 small animal reporter, submission 1
SCGE ID:1076 - Submission Date: 2022-10-19 
Experiment Name Type Description
Validating Gene Editing Reporter 12 (GER12) Mouse Model in Heterozygous Blastocysts

In Vitro WT female mouse and homozygous male mouse containing the Gene Editing Reporter 12 (GER12) reporter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Cas9 RNP along with test guides and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for Katushka2S (RFP) fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 58-69. Total blast analyzed by Sanger sequencing, 51-52.
NEW MODEL DEVELOPMENT - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - TLR7 small animal reporter, submission 1
SCGE ID:1077 - Submission Date: 2022-10-19 
Experiment Name Type Description
Validating Traffic Light Reporter 7 (TLR7) Mouse Model for NHEJ in Heterozygous Blastocysts

In Vitro WT female mouse and homozygous male mouse containing the Traffic Light Reporter 7 (TLR7) reporter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Cas9 RNP along with test guides and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for Katushka2S (RFP) fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 169. Total blast analyzed by Sanger sequencing, 117.
Validating Traffic Light Reporter 7 (TLR7) Mouse Model for HDR in Heterozygous Blastocysts

In Vitro WT female mouse and homozygous male mouse containing the Traffic Light Reporter 7 (TLR7) reporter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Cas9 RNP along with test guides and HDR donor template, and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for Katushka2S (RFP) fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 103. Total blast analyzed by Sanger sequencing, 64.
NEW MODEL DEVELOPMENT - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - GER14 small animal reporter, submission 1
SCGE ID:1078 - Submission Date: 2022-10-19 
Experiment Name Type Description
Validating Gene Editing Reporter 14 (GER14) Mouse Model in Heterozygous Blastocysts

In Vitro WT female mouse and homozygous male mouse containing the Gene Editing Reporter 14 (GER14) reporter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with various CRISPR/Cas9 combinations (or Cre) and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for Katushka2S (RFP) fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 46-164. Total blast analyzed by Sanger sequencing, 39-142.
NEW MODEL DEVELOPMENT - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - GER18 small animal reporter, submission 1
SCGE ID:1079 - Submission Date: 2022-10-19 
Experiment Name Type Description
Validating Gene Editing Reporter 18 (GER18) Mouse Model in Heterozygous Blastocysts

In Vitro WT female mouse and homozygous male mouse containing the Gene Editing Reporter 18 (GER18) reporter driven by the EF-1 alpha promoter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Cas9 RNP along with two guides and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for GFP fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced to determine efficiency of deleting the IVS2-654 splicing variant. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 61. Total blast analyzed by Sanger sequencing, 54.
NEW MODEL DEVELOPMENT - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - GER19 small animal reporter, submission 1
SCGE ID:1080 - Submission Date: 2022-10-19 
Experiment Name Type Description
Validating Gene Editing Reporter 19 (GER19) Mouse Model in Heterozygous Blastocysts

In Vitro WT female mouse and homozygous male mouse containing the Gene Editing Reporter 19 (GER19) reporter driven by the CAG promoter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with Cas9 RNP along with two guides and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for GFP fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced to determine efficiency of deleting the IVS2-654 splicing variant. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 53. Total blast analyzed by Sanger sequencing, 48.
VALIDATION - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - SATC-Chen, Submission 2 (On- and Off-target data)
SCGE ID:1082 - Submission Date: 2022-12-13 
Experiment Name Type Description
Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear

In Vivo Delivery of CRISPR/Cas9 via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice. Subset of mice were administered LNP via canalostomy injection compared to uninjected control mice. Tissues were harvested 6 days after LNP administeration. On-target and off-target editing was assessed.
Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear

In Vivo Repeat experiment of CRISPR/Cas9 delivery via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice. Subset of mice were administered LNP via canalostomy injection. Control mice were admininstered a blank LNP. Tissues were harvested 6 days after LNP administration. On-target editing was assessed by RFP (tdTomato) signal.
VALIDATION - The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - SATC-Gong, Submission 2 (On- and Off-target data)
SCGE ID:1083 - Submission Date: 2022-12-13 
Experiment Name Type Description
Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain

In Vivo Delivery of CRISPR/Cas9 via ribonuclear protein (RNP) loaded nanocages (NC) to the brain in Ai14 mice by intracranial bilateral injection. Tissues were harvested 14 days after NC administeration. On-target and off-target editing was assessed.