Description:
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GE-Low PPO Avocado lines were developed using the CRISPR/Cas9 technology delivered into plant cells by the Ribonucleoprotein (RNP) method.
The intended phenotype of GE-Low PPO Avocado lines is reduced levels of polyphenol oxidase proteins, enzymes responsible for enzymatic darkening and discoloration in avocado fruit flesh (mesocarp tissue). When avocados are mashed or cut, PPO enzymes are released by the disruption of the cell structure, resulting in unappealing discoloration.
CRISPR/Cas9 was used to generate one or more targeted DNA double-stranded breaks within the coding region of at least one PPO gene. No external repair template was provided. Instead, the plant’s own cellular mechanism repairs the breaks, resulting in small nucleotide deletions, additions, or substitutions at the break sites on each allele which likely are repaired in a slightly different way.
The final GE-Low PPO Avocado lines were molecularly screened for double-allele knockouts. With the RNP method, the final selected lines do not contain any introduced DNA from the CRISPR/Cas9 system.
Source:
- USDA-APHIS letter of inquiry 20-168-35_air
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Links regarding to this GMO:
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USDA APHIS https://www.aphis.usda.gov/biotechnology/downloads/reg_loi/20-168-35_air_inquiry_a2_cbidel.pdf
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Transformation / Modification technique:
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CRISPR/Cas
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