Model for simulating ALS constructed based on CASP4 and its construction method

A method for constructing an amyotrophic lateral sclerosis (ALS)-simulating model based on a caspase-4 (CASP4) gene, comprising the following steps:

• (1) constructing a targeting fragment CAG-loxP-stop-loxP (LSL)-human CASP4(hCASP4)-posttranscriptional regulatory element of woodchuck hepatitis virus (WPRE)-polyA for a knock-in of the CASP4 gene;
• (2) injecting a gRNA, a Cas9 mRNA, and the targeting fragment CAG-LSL-hCASP4-WPRE-polyA into a mouse zygote, culturing, and passaging to produce a hCASP4flox mouse with the CASP4 gene stably inherited;
• and
• (3) crossing the hCASP4flox mouse with a Nestin-Cre driver mouse to produce a double-positive heterozygous mouse, namely, a mouse model, wherein in the mouse model, the CASP4 gene is specifically expressed in a nervous system;
• wherein amplification primers for identifying the double-positive heterozygous mouse are as follows:
• Caspase-4-F-C1:
• 5′-TCTACCTCTTTCCTGGCAATGACTACA-3′,
• as shown in SEQ ID NO:
• 2;
• Caspase-4-R-C1:
• 5′-CTTTATTAGCCAGAAGTCAGATGC-3′,
• as shown in SEQ ID NO:
• 3;
• Caspase-4-F-C2:
• 5′-CACTTGCTCTCCCAAAGTCGCTC-3′,
• as shown in SEQ ID NO:
• 4;
• Caspase-4-R-C2:
• 5′-ATACTCCGAGGCGGATCACAA-3′,
• as shown in SEQ ID NO:
• 5;
• Nestin-F-N1:
• 5′-CCTTCCTGAAGCAGTAGAGCA-3′,
• as shown in SEQ ID NO:
• 6;
• Nestin-R-N:
• 5′-GCCTTATTGTGGAAGGACTG-3′,
• as shown in SEQ ID NO:
• 7;
• and
• Nestin-F-N2:
• 5′-TTGCTAAAGCGCTACATAGGA-3′,
• as shown in SEQ ID NO:
• 8.