Additional CRISPR/Cas9 expression vector and construction method and application thereof
A technology of expression vector and construction method, which is applied in the field of cell engineering, can solve the problem of destroying the sequence in the genome, and achieve the effect of avoiding potential safety hazards
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Embodiment 1
[0044] Example 1. Episomal CRISPR / Cas9 expression vector.
[0045] The episomal CRISPR / Cas9 expression vector described in this embodiment refers to a CRISPR / Cas9 expression vector containing the S / MAR sequence. The S / MAR sequence has nucleotides as shown in SEQ ID NO:1; The / MAR sequence is located between the coding region sequence capable of encoding the protein and the bGH poly(A) sequence in the vector.
Embodiment 2
[0046] Example 2. Episomal CRISPR / Cas9 expression vector.
[0047] The episomal CRISPR / Cas9 expression vector described in this embodiment refers to a CRISPR / Cas9 expression vector containing an S / MAR sequence. The S / MAR sequence has nucleotides as shown in SEQ ID NO:1. The / Cas9 expression vector contains the EGFP coding region sequence, such as the EGFP coding region sequence from the px330-EGFP vector. The S / MAR sequence is located between the EGFP coding region sequence and the bGH poly(A) sequence in the CRISPR / Cas9 expression vector between.
Embodiment 3
[0048] Example 3. Episomal CRISPR / Cas9 expression vector.
[0049] The episomal CRISPR / Cas9 expression vector described in this embodiment refers to a CRISPR / Cas9 expression vector containing the S / MAR sequence. The S / MAR sequence has nucleotides as shown in SEQ ID NO:1; The / MAR sequence is located between the coding region sequence capable of encoding the protein and the bGH poly(A) sequence in the vector. The sgRNA sequence in the CRISPR / Cas9 expression vector is one of SEQ ID NO: 2-4.
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