CUSTOM MODEL GENERATION SERVICES

CRISPR-Cas9? ???? ??? ??? ???? ??

????? CRISPR-Cas9 ??? ???? ?? 4,000? ??? ??? ??? ??? ???? ???, ??? ?? ??? ?? ?? ?????. CRISPR-Cas9 ??? ??? ?? ?? ???? ??? ?? ???, ???(KO) ???, ??? ???(cKO) ???, ??(KI) ??? ??? ??? ? ????.

CRISPR-Cas9 ??? ALEN, ZFN, ES? ?? ?? ??? ?? ???? ???? ??? ?????.

Advantages:

???: ??? ??(random insertion)?? ??? ???(no serial recombination)? ?? ??? ??
???: ????? ???? ?? ???? ???? ??? ?? ??? ??
Multi-modification: ?? ?? loci, gene, large fragment? ??? ? ? ??
??? ??? ??: C57BL/6J, C57BL/6N, BALB/c, FVB, NOD/LTJ, NCG ? ??? ??? ???? ??? ?? ?? ??

?????? ?? ??? ??? ?? ??? ??? ?? ??

??????(Embryonic Stem Cell, ES) ?? ???(Homologous Recombination, HR) ??? ?? DNA? ES?? ??? ?? ???? ??? ?? ?????. ??? ??? ?? ??? ????? ??? DNA? ??? CRISPR-Cas9 ??? ??? ??? ???? ??? ??? ?? ??? ?????.

??:

?? ???? ??? ??(Self-deletion of resistance genes)?? ?? ?? ??? 3?? ??
BAC ?? ??? 100-300kb? ??? ??(knock-in)? ? ??
2005??? ??? ? ??? ???, 700? ??? ??? ? ??? ???? ????? ?????
129S6/SvEvTac ? C57BL/6N ? ??? ??? ???? ??? ??? ? ??

????? ??? ??? ??? ??? ?? ??

????? ??? ?? ???(overexpressed) ?? ??? ???(conditionally overexpressed) ???? ??? ??? ???? ?? ?????. ????? ??? ??? ??? ?? ?? ?, ????(microinjection) ??? DNA? ??? ??? ??(pronucleus)? ?? ????? DNA? ??? ???? ???? ??(random insertion)???. ??(KI) ??? ???? ??, ??? ??? ?? ?? ? ???? ??? H11 ?? Rosa26 site? ???? ?????.

Comparison of transgenic technology and KI at the H11 or Rosa26 site
Transgenic	H11/Rosa26 Knockin
F0 generation in as little as 8 weeks	F1 generation in as little as 16 weeks
Low technical difficulty	High technical difficulty
Different expression levels between founder lines, and expression levels over time (typically expression levels decrease)	Same expression levels in all founder lines, and constant expression levels over time
Random insertion site and number of copies	Accurate insertion location and gene copy number by design
Easy to alter or destroy the expression of other endogenous genes	Does not affect expression other endogenous genes
Poor genetic stability	High genetic stability
Large-scale work in establishing a stable line	Produces a stable line directly for analysis

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