Part:BBa_K5167670

CRISPR/Cas9 target site sequence of OsGlb

It is a CRISPR/Cas9 target site sequence in the OsGlb gene of rice.OsGlb is a single gene encoding the allergen 19kDa globulin in rice endosperm. It is located on chromosome 5: 24576939-24577725 reverse strand. Its access nomuber is os05g0499100. It has only one exonic region.

Usage and Biology

When the sgRNA in crispr/cas9 gene editing system guides cas9 protein to target the globulin coding gene OsGlb in the rice genome, this element is the recognition sequence of sgRNA, and the sgRNA for OsGlb needs to be designed according to the sequence of this element. When Cas9 protein recognizes and cleaves the DNA duplex of the target site sequence under the guidance of the sgRNA corresponding to OsGlb, a gap will be formed on the duplex to realize the mutation of OsGlb gene.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

This is the target site sequence of the globulin coding gene OsGlb in the rice genome targeted by CRISPR/Cas9 gene editing system. It is located in the CDS region, adjacent to the PAM, and upstream of the PAM. After the Cas9 protein is guided by sgRNA to cleave the target site sequence, a base a is inserted between the penultimate and fourth bases of the target site sequence. It leads to a frameshift mutation in OsGlb, and this frameshift mutation leads to the premature appearance of a codon reading frame at the stop codon. In this case, the globulin encoded by OsGlb is hardly expressed. The size of the mutant globulin molecule is slightly smaller, the hydrophobicity and instability are increased, and the binding ability with human immunoglobulin E is greatly reduced.

Functional Parameters

protein	-NA-
target	-NA-

This sgRNA target site sequence is located in CDS, and its PAM is CGG. Its CG content is 12 / 20.