BioVector? NTCC? LbABE8e plasmid基因編輯質(zhì)粒BioVector?載體

The BioVector? LbABE8e plasmid (available as BioVector? plasmid #138504 and related variants) is a key tool in adenine base editing, facilitating efficient A?T-to-G?C conversions in mammalian cells. Plasmid Details Plasmid Name: LbABE8e Plasmid #: BioVector?138504 Deposited by: David Liu Lab Vector Backbone: pCMV with a pBR322 origin Vector Type: Mammalian expression Bacterial Resistance: Ampicillin (100 μg/mL) Bacterial Growth Strain: DH5alpha is a suggested strain for propagation. General Function: Adenine base editor for A-to-G base conversion. Key Components The LbABE8e plasmid is an expression vector that fuses two primary functional elements: an engineered deaminase and a deactivated CRISPR-associated protein. TadA-8e Deaminase Variant: The deaminase domain has been highly evolved (hence the "8e" designation) to improve activity and compatibility with various Cas proteins. It converts adenine (A) to inosine (I), which is then read as guanine (G) during DNA replication. dLbCas12a (dead Lachnospira bacterium Cas12a): This nuclease-dead Cas12a protein (containing mutations to abolish DNA-cutting activity) is responsible for guiding the base editor to the specific target DNA sequence. Guide RNA (gRNA): A separate guide RNA is required to direct the LbABE8e protein to the desired genomic location. Promoter: A CMV promoter drives high expression of the fusion protein in mammalian cells. Origin of Replication (ori): The pBR322 origin facilitates replication of the plasmid within bacteria for propagation. Antibiotic Resistance Gene: An ampicillin resistance gene allows for selection of bacteria that have successfully taken up the plasmid. Usage and Maintenance Host Organism: This is a mammalian expression vector. Propagation in Bacteria: Grow in LB media supplemented with 100 μg/mL ampicillin. Reference: This system was originally described in Nature Biotechnology.

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