Conditional Colon Tumor Induction in Mice

All experiments were performed in accordance with the Memorial Sloan Kettering Institutional Animal Care and Use Committee (IACUC) under protocol number 11‐06‐012. Doxycycline was administered via food pellets (625 mg·kg⁻¹) (Harlan Teklad). 4‐Hydroxytamoxifen (4OHT, Sigma Aldrich, St. Louis, MO, USA, 70% Z‐isomer) was delivered by a single intraperitoneal injection (0.5 mg/mouse) at 5–6 weeks of age. LSL‐Kras (B6.129S4‐Kras^tm4Tyj/J) and Lgr5‐CreER (B6.129P2‐Lgr5^{tm1(cre/ERT2)Cle}/J) animals were purchased from Jackson Laboratories. CAGs‐LSL‐rtTA3 (B6. Cg‐Gt(ROSA)26Sor^{tm1(CAG‐rtTA3)Slowe}/LdowJ) and TG‐Apc.3374 (Col1a1^{tm4(tetO‐GFP/RNAi:Apc)Slowe}) mice were described previously [27 (link), 28 (link)]. We used a GEMM in which the Apc gene can be conditionally suppressed using a doxycycline‐regulated shRNA to develop colon tumors (shApc/Kras^wt mice), as described previously [27 (link), 28 (link)]. A KRAS‐mt line was developed by crossing conditional KRAS‐mt allele‐carrying mice (LSL‐Kras^G12D) with the shApc mice to develop shApc/Kras^G12D mice. Mouse colonic organoids were isolated as previously published [28 (link)].

Free full text: Click here

Kim J.K., Marco M.R., Choi S., Qu X., Chen C., Elkabets M., Fairchild L., Chow O., Barriga F.M., Dow L.E., O’Rourke K., Szeglin B., Yarilin D., Fujisawa S., Manova‐Todorova K., Paty P.B., Shia J., Leslie C., Joshua Smith J., Lowe S., Pelossof R., Sanchez‐Vega F, & Garcia‐Aguilar J. (2021). KRAS mutant rectal cancer cells interact with surrounding fibroblasts to deplete the extracellular matrix. Molecular Oncology, 15(10), 2766-2781.

Publication 2021

food pellets 4‐Hydroxytamoxifen (4OHT,

4 hydroxytamoxifen Allele Animals Apc gene Colon tumors Colonic Doxycycline Ert2 Food Institutional animal care and use committee Intraperitoneal injection Isomer Kras Mouse Organoids Pellets Rnai Rosa Shrna

Corresponding Organization : Memorial Sloan Kettering Cancer Center

Other organizations : Ben-Gurion University of the Negev, Cornell University, Howard Hughes Medical Institute

Top 5 similar protocols

Variable analysis

independent variables

Doxycycline administration via food pellets (625 mg·kg−1)
4-Hydroxytamoxifen (4OHT) delivery by a single intraperitoneal injection (0.5 mg/mouse) at 5–6 weeks of age

dependent variables

Development of colon tumors in the shApc/Kras^wt and shApc/Kras^G12D mouse models

control variables

Mouse models used: LSL-Kras, Lgr5-CreER, CAGs-LSL-rtTA3, and TG-Apc.3374

controls

Positive control: shApc/Kras^wt mouse model to develop colon tumors
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!