Table 2 Twenty-six PgABCA2 cDNA mutations in 11 Cry2Ab-resistant pink bollworm larvae from the CRISPR-R2 strain (n = 5) and CRISPR-R2 X APHIS-S F₂ progeny (n = 6).

From: CRISPR-mediated mutations in the ABC transporter gene ABCA2 confer pink bollworm resistance to Bt toxin Cry2Ab

cDNA mutation^a	Larva.clone^b	Codon^c	Exon^d	Type^e	Effect^f	sgRNA sites^g	Current source^h	Previous sourceⁱ
c.295_559delins	3.1	99	2	fs	Stop	1	CRISPR-R2	No
c.677_3096delins	3.1	226	3–17	fs	Stop	2	CRISPR-R2	No
c.240_3100del	3.2	80	2–17	fs	Stop	No (1–5)	CRISPR-R2	No
c.295_3100delins	3.3	99	2–17	fs	Stop	1	CRISPR-R2	No
c.240_3096del	3.15	80	2–17	fs	Stop	No (1–5)	CRISPR-R2	No
c.3313_3589del	5A.7, A.1.1, A.2.2, A.2.9, 10.29	1105	19–20	fs	Stop	No	Both	Bt4-R2
c.3097_3100del	5A.11, C.4.6	1033	18	fs	Stop	No	Both	BX-R
c.3418_3589del	5A.11, 5A.12, 10.5, 10.11, C.3.4	1140	20	fs	Stop	No	Both	Bt4-R2
c.241_1409del	5B.4, 5B.6, 5B.8	80	2–7	fs	Stop	No (1–4)	CRISPR-R2	No
c.1835_1976del	5B.8	612	11	fs	Stop	No	CRISPR-R2	No
c.2125_2131del	5B.4, 5B.6, 5B.8, C.4.6, C.4.8, C.4.10	708	12	fs	Stop	5	Both	No
c.294_2128del	9A.8, 9A.9, 9A.10, 9A.13, 9B.12, 9B.14, 9B.16	98	2–12	fs	Stop	1, 5	CRISPR-R2	No
c.2220_2221delinsAG	9A.8	740	12	fs	Stop	No	CRISPR-R2	No
c.4438_4439ins	9A.10	1479	26–27	fs	Stop	No	CRISPR-R2	No
c.3589_3714del	9B.12	1196	21	InF	In-frame del	No	CRISPR-R2	No
c.3555_3590ins	9B.14	1185	20	fs	Stop	No	CRISPR-R2	No
c.1021_4668del	10.1, 10.21, 10.22	340	5–28	InF	In-frame del	No (4–5)	CRISPR-R2	No
c.1090_1234del	10.5, 10.11, A.2.9, C.3.4, C.4.6	364	6	fs	Stop	No	Both	Bt4-R2, BX-R, India
c.1755_4587del	10.19	584	10–29	fs	Stop	No (5)	CRISPR-R2	No
c.559_4884del	10.24	186	3–29	InF	In-frame del	No (2–5)	CRISPR-R2	No
c.3556_3588delins	18.6, 18.7, 18.20, C.3.6	1185	20	fs	Stop	No	CRISPR-R2	Bt4-R2
c.4598_4599ins	C.3.6	533	27	fs	Stop	No	F₂	No
c.294_295delinsGAGA	C.4.6, C.4.8, C.4.10	98	2	fs	Stop	1	F₂	No
c.674_681del	C.4.6, C.4.8, C.4.10	225	3	fs	Stop	2	F₂	No
c.283_296del	J.5.2, J.5.3, J.5.6, J.6.4	95	2	fs	Stop	1	F₂	No
c.679_2229delins	J.5.2, J.5.3, J.5.6, J.6.4	227	3–12	fs	Stop	2	F₂	No

^acDNA nomenclature showing the nucleic acid sequence changes in PgABCA2 (MG637361.1) based on the recommendations by the Human Genome Variation Society (http://www.hgvs.org/).
^bEntries start with a number for the five larvae from the CRISPR-R2 strain (3, 5, 9, 10 and 18) and with a letter for the six larvae from F₂ progeny of three single-pair families from CRISPR-R2 X APHIS-S crosses (A, C and J). For each entry, the first number indicates the larva and the second number indicates the clone (e.g., 3.1 indicates larva 3, clone 1). For 5A and 5B as well as 9A and 9B, two distinct PCR products (A and B) were cloned and sequenced from a single larva.
^cCodon number beginning from the initiation codon and indicating disrupted position in the coding sequence.
^dDisrupted exon in the coding sequence.
^eType of mutation (fs, frame shift; InF, in-frame mutation).
^fResult of mutation in the coding sequence (Introduction of premature stop codon; In-frame del; In-frame deletion).
^gNumbers 1, 2 or 5 indicate the sgRNA site directly affected; no indicates none of the five sgRNA sites were directly affected and numbers in parentheses indicate sgRNA sites indirectly affected (mutations occurred in exons containing sgRNA sites).
^hSource of larvae in the current study: CRISPR-R2, F₂ progeny of single-pair families from CRISPR-R2 X APHIS-S, or both.
ⁱNo indicates not found in previous studies; other entries indicate the same cDNA mutation was previously found in one or more of the following sets of Cry2Ab-resistant insects: lab-selected Bt4-R2 and BX-R strains from Arizona and field-selected populations from India^29,30.

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Table 2 Twenty-six PgABCA2 cDNA mutations in 11 Cry2Ab-resistant pink bollworm larvae from the CRISPR-R2 strain (n = 5) and CRISPR-R2 X APHIS-S F₂ progeny (n = 6).

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